EP2622338A1 - Tube capillaire pour l'électrophorèse - Google Patents

Tube capillaire pour l'électrophorèse

Info

Publication number
EP2622338A1
EP2622338A1 EP11764145.6A EP11764145A EP2622338A1 EP 2622338 A1 EP2622338 A1 EP 2622338A1 EP 11764145 A EP11764145 A EP 11764145A EP 2622338 A1 EP2622338 A1 EP 2622338A1
Authority
EP
European Patent Office
Prior art keywords
capillary tube
plastic
hole
capillary
μιη
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.)
Withdrawn
Application number
EP11764145.6A
Other languages
German (de)
English (en)
Inventor
Dominik Mueller
Karsten Pinkwart
Jens Tuebke
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Original Assignee
Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV filed Critical Fraunhofer Gesellschaft zur Forderung der Angewandten Forschung eV
Publication of EP2622338A1 publication Critical patent/EP2622338A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44756—Apparatus specially adapted therefor
    • G01N27/44791—Microapparatus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G01N27/44704—Details; Accessories
    • G01N27/44713—Particularly adapted electric power supply
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
    • G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/416—Systems
    • G01N27/447—Systems using electrophoresis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2223/00—Investigating materials by wave or particle radiation
    • G01N2223/60—Specific applications or type of materials
    • G01N2223/628—Specific applications or type of materials tubes, pipes

Definitions

  • the present invention relates to a capillary for capillary electrophoresis and a chemical separation and analysis device containing this capillary.
  • the electrophoretic separation is caused by the application of a voltage.
  • Charged particles migrate in the electric field to the corresponding poles, e.g. negatively charged particles to the positive pole.
  • Each charged particle shows different speeds in the electric field due to different mobilities.
  • the mobility depends on the charge number, the radius and the forming hydrate layer of the particle.
  • EEF electroosmotic flow
  • Optical detectors can thus be used "on-column" without coming into contact with the live fluid, but only compounds that absorb in the appropriate wavelength range can be detected, eg, mono- and oligosaccharides have no chromophore
  • These devices must be used with other sensitive detectors, such as mass selective and electrochemical detectors, but these detectors come into contact with the live fluid, resulting in significant degradation or failure of these detectors Lead out separating line, before the fluid reaches the detector.
  • the fluid passes through an interface to the mass spectrometer, where a liquid (sheath liquid) is fed into the interface in order to be able to remove the voltage from the capillary, an undesirable effect of which is the dilution with the fluid from the capillary, thereby significantly reducing the detection sensitivity.
  • a liquid sheath liquid
  • the glass separation channel is not coated, which results in a high EOF.
  • This difference in the EOF leads to an indirect hydrodynamic flow after branching to the end of the canal. This flow is assisted by increasing the flow resistance in the side arm by making the length of the side arm longer than the distance from the branch to the end of the channel.
  • Electrochemical detection is referred to as "off-channel detection” when the voltage is to be taken out prior to electrochemical detection.
  • a “decoupler” is described in the above publication and by H. Chen et al. in Trends in Analytical Chemistry, Vol. 26, No. 2, 2007, several variants are presented. J.S. Rossier et al, Journal of Electroanalytical Chemistry, 492 (2000), 15, describes a construction in which microholes are integrated at the end of the separation channel, which are made of a different polymer material. This construction allows the voltage before electrochemical detection from the separation channel
  • the invention consists in the provision of a suitable chemical separation and analysis device containing the capillary according to the invention and a chemical separation and analysis method using the
  • this object is achieved by providing a plastic capillary tube for the
  • the plastic capillary tube has an inlet and an outlet opening and further comprises at least one hole in the Kapillarrschreibchenwand and the diameter of the hole on the inside of the Kapillarrschreibchenwand thin ) in the range of 0.5 ⁇ to 30 ⁇ .
  • the capillary tube of the invention is a polymer capillary tube, i. one out of one
  • Plastic capillary tubes show less surface adsorption than conventional glass capillary tubes for compounds that tend to have a strong tendency due to their chemical structure. Examples are proteins and oligosaccharides. For such classes of compounds, the electrophoretic separation in plastic capillary tubes has a favorable effect. Furthermore, chemically and mechanically stable plastic capillary tubes can be produced more cheaply. As discussed above, the use of sensitive and selective
  • the plastic capillary tube has at least one hole in the capillary tube wall and the diameter of the hole on the inside of the capillary tube wall thin) in the range of 0.5 ⁇ to 30 ⁇ . Due to the selected diameter of the hole in the capillary wall, it is made possible on the one hand to bring out the tension, but on the other hand leakage of the fluid from the hole in the capillary tube wall is prevented as far as possible under given experimental conditions.
  • the diameter of the hole is located on the inside of the hole
  • Capillary tube wall thin in the range of 1 ⁇ to 20 ⁇ , more preferably 2 ⁇ to 12 ⁇ .
  • the determination of the hole diameter may e.g. carried out via optical evaluation of light microscopy images.
  • the hole or bore diameter is determined manually by inserting circles via the 3-point method or by determining the center point and radius in the light microscope image. The hole must be positioned in the middle.
  • the diameter of the hole on the inside of the capillary tube wall is thin) smaller than the diameter of the hole on the outside of the capillary tube wall dL (outer).
  • a hole whose cross-sectional opening is not circular eg.
  • the hole has a conical or conical shape.
  • a conical shape of the hole can be e.g. by laser drilling, i.e., "burning in” a hole in the capillary wall by laser action.
  • the ratio of the hole diameter at the inside of the hole diameter is the ratio of the hole diameter at the inside of the hole diameter
  • Capillary tube wall dL (inside) dL (outside) in the range of 1/2 to 1/7, more preferably 1/3 to 1/5.
  • the ratio dL (inside) / dL (outside) can be determined by means of a lateral
  • the axis of the hole extends at an angle in the range of 90 ° ⁇ 20 °, more preferably in the range of 90 ° ⁇ 10 ° to the longitudinal axis of the
  • the plastic capillary tube of the present invention may have only one or alternatively two or more holes in the capillary tube wall.
  • plastic capillary tube only one hole in the
  • Capillary tube wall has.
  • the plastic capillary tube has further holes in the capillary tube wall, with respect to the properties of these additional holes, reference may be made to the above statements regarding the first hole. If the plastic capillary tube has two holes in the capillary tube wall, it is preferable for the two holes in the capillary tube wall to be as close as possible to each other, ie, to be on the same axis or uniaxial.
  • Plastic capillary tube for capillary electrophoresis can be varied accordingly.
  • the outer diameter of the plastic capillary tube is outside ) in the range of 50 ⁇ to 200 ⁇ , more preferably from 75 ⁇ to 170 ⁇ and the
  • Inner diameter of the plastic capillary tube inside is preferably in the range of 10 ⁇ to 150 ⁇ , more preferably from 50 ⁇ to 125 ⁇ .
  • the thickness of the Kapillarrschreibchenwand in the range of 1 ⁇ to 25 ⁇ .
  • the total length L 0 of the Kunststoffkapillarrschreibchens may vary depending on the type of sample to be analyzed and the required separation conditions.
  • a suitable total length L 0 of the plastic capillary tube may be, for example, in the range of 40 cm to 150 cm, more preferably 55 cm to 100 cm.
  • Plastic capillary tubes of such dimensions are commercially available.
  • the hole having a diameter which is in the above-indicated diameter range can be mounted in the capillary tube wall by methods generally known to those skilled in the art. Exemplary in this
  • the laser drilling for example, a microchip laser with a wavelength of 532 nm can be used.
  • the spot irradiation technique is used.
  • the laser beam is focused in this way that a focal point arises on the capillary surface.
  • the desired inner diameter of the hole can ultimately be determined.
  • the capillary tube at its respective ends, ie the capillary tube has at one end an inlet opening for receiving the sample liquid and at the other end an outlet opening.
  • the hole in the capillary tube wall is as close as possible to the outlet opening of the tube.
  • the hole is at a distance Li from the end of the capillary tube having the outlet opening and the total length of the plastic capillary tube is L 0 and the ratio Li / L 0 is in the range of 1/8 to 1/500, more preferably 1 / 20 to 1/100.
  • Plastic capillary tube can be made, e.g. polyimide,
  • the present invention provides a chemical separation and analysis apparatus comprising
  • the capillary electrophoresis unit comprises a receptacle for a sample or electrolyte liquid in the region of the end of the capillary tube having the inlet opening, so that the liquid can easily enter the capillary tube
  • Capillary tube can be injected, e.g. by hydrodynamic or electrokinetic injection.
  • an electrode E2 is mounted in the region of the receptacle so that it can be immersed in the sample or electrolyte liquid when the receptacle is filled or can be in electrically conductive contact therewith.
  • the capillary electrophoresis unit comprises a device element for establishing an external pressure in the plastic capillary tube.
  • a device element for establishing an external pressure in the plastic capillary tube.
  • the liquid is conducted into the capillary at a different flow rate.
  • a liquid pump is preferably positioned in front of the voltage inlet.
  • the chemical separation and comprises
  • the detection unit is preferably an electrochemical detector, mass-selective detector, conductivity detector, AOW or SAW sensors (AOW: surface acoustic waves or SAW: surface acoustic
  • Impendanzspektroskopiesensor or impedance sensor provides optically based detectors, such as UV, VIS, fluorescence, and refractive index detectors, or combinations of these detectors.
  • optically based detectors such as UV, VIS, fluorescence, and refractive index detectors, or combinations of these detectors.
  • the present invention provides a chemical separation and analysis method comprising introducing one
  • Plastic capillary tubes may be made in a conventional manner known to those skilled in the art, e.g. by hydrodynamically or electrokinetically performed injection.
  • Buffer liquid is, in which an electrode El dips or which is in electrically conductive contact with an electrode El.
  • Buffer liquid is in contact. In this container can then be embedded in the electrode El.
  • Another electrode E2 preferably dips into a
  • Sample liquid or is in electrically conductive contact with this, wherein the sample liquid is present in a vessel, preferably in the field of
  • Inlet opening of the capillary tube is positioned.
  • the voltage across the hole in the tube wall is effectively led out of the capillary tube, so that the sample liquid after passing through the hole is no longer under tension and thus can be fed to a downstream detection unit.
  • the sample liquid introduced into the plastic capillary tube is subjected to a pressure in the range of 1 to 500 mbar, more preferably 5 mbar to 150 mbar.
  • a pressure in the range of 1 to 500 mbar, more preferably 5 mbar to 150 mbar.
  • the sample liquid is supplied after passing the hole in the Kapillarrschreibchenwand a downstream detection unit.
  • the detection unit is preferably an electrochemical detector, mass-selective detector, conductivity detector, AOW or SAW sensors (AOW: surface acoustic waves or SAW: surface acoustic waves), impedance spectroscopy sensor or impedance sensor, optically based Detectors, such as UV, VIS, fluorescence, and refractive index detectors, or combinations of these detectors.
  • Figure 1 shows schematically a preferred embodiment, which will be described in more detail below.
  • the preferred chemical separation and analysis apparatus shown in Figure 1 includes the plastic capillary tube 1 described above having a hole 2 in the capillary tube wall.
  • the inlet opening having the end of
  • Sample liquid 3 a In this liquid, an electrode 4 is also attached.
  • a buffer vessel 5 is placed at the hole 2 in the capillary tube wall.
  • the second electrode 6 In this buffer or electrolyte vessel 5, the second electrode 6 is attached.
  • a DC voltage is applied between the electrodes (voltage between 1 and 30kV). The voltage is passed through the plastic capillary tube 1 between the capillary inlet opening and the hole 2 in the capillary tube wall and closes the circuit. In this capillary section electrophoretic separation takes place.
  • the first buffer or electrolyte vessel is exchanged with the sample vessel 3. The injection is carried out hydrodynamically or electrokinetically.
  • the polarity of the electrodes 4, 6 can be switched to negative-positive or vice versa, depending on the analytical question. If negatively charged analytes are to be detected, the electrode in the first buffer or electrolyte vessel 3 is contacted negatively and positive in the second buffer or electrolyte vessel 5. In the electrophoretic separation section, the negatively charged analytes migrate to the positive pole and are separated due to their different mobility. A second electrophoretic effect is the electroosmotic flow EOF already discussed above, which transports the entire bulk part in the capillary tube in the direction of the positive pole. As a result, negatively charged analytes are accelerated and positively charged analytes are slowed down by the capillary tube 1. Neutral particles, on the other hand, migrate with the electroosmotic flow. Since, when using the capillary tube 1 according to the invention, the voltage is led out through the hole 2 in the capillary tube wall, there is no space after the hole 2
  • a commercially available polyimide capillary tube having an outer diameter of 168 ⁇ m and an inner diameter of 122 ⁇ m was used. The wall thickness was therefore 23 ⁇ .
  • Such a plastic capillary tube is commercially available, e.g. from Goodfellow.
  • the capillary tube had a total length of 60 cm. At a distance of 7 cm from the end of the capillary having the outlet opening were two opposite holes in the
  • the detection unit used was a UV detector from Dionex with a flow cell, which was connected to the plastic capillary tube via a transfer line.
  • a commercially available aqueous borate solution was used as a buffer.
  • Nitrate was used as the UV-active test substance.
  • a voltage of U 15 kV was applied. Furthermore, an external pressure of 4 psi was applied. At the point of the two holes in the
  • Capillary tube wall flows a current of 55 ⁇ . In other places of the capillary, the flow of current is interrupted. Further, despite an external pressure of 4 psi, there is no drop formation due to leaking buffer liquid from the holes in the capillary wall. The example shows that stress can escape at the location of the holes in the capillary wall, but at a given pressure, the buffer liquid is retained in the capillary tube.
  • inventive chemical separation and analysis device realize, inter alia, the following advantages:
  • Detection sensitivity is maintained, no sheath fluid is used, and the sample (injection volume) is not diluted.

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  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Electrochemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Engineering & Computer Science (AREA)
  • Power Engineering (AREA)
  • Dispersion Chemistry (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
  • Optical Measuring Cells (AREA)
  • Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
  • Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
  • Electrostatic Separation (AREA)

Abstract

La présente invention concerne un tube capillaire en matière plastique destiné à l'électrophorèse capillaire, le tube capillaire en matière plastique présentant une ouverture d'entrée et une ouverture de sortie, et au moins un orifice étant pratiqué dans la paroi du tube capillaire, orifice dont le diamètre, sur le côté intérieur de la paroi de tube capillaire dL(intérieur) vaut de 0,5 µm à 30 µm.
EP11764145.6A 2010-09-27 2011-09-23 Tube capillaire pour l'électrophorèse Withdrawn EP2622338A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE102010041433A DE102010041433A1 (de) 2010-09-27 2010-09-27 Kapillarröhrchen für die Elektrophorese
PCT/EP2011/066551 WO2012041767A1 (fr) 2010-09-27 2011-09-23 Tube capillaire pour l'électrophorèse

Publications (1)

Publication Number Publication Date
EP2622338A1 true EP2622338A1 (fr) 2013-08-07

Family

ID=44735901

Family Applications (1)

Application Number Title Priority Date Filing Date
EP11764145.6A Withdrawn EP2622338A1 (fr) 2010-09-27 2011-09-23 Tube capillaire pour l'électrophorèse

Country Status (7)

Country Link
US (1) US9410925B2 (fr)
EP (1) EP2622338A1 (fr)
JP (1) JP5685651B2 (fr)
KR (1) KR101516098B1 (fr)
CA (1) CA2811109A1 (fr)
DE (1) DE102010041433A1 (fr)
WO (1) WO2012041767A1 (fr)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US11899294B2 (en) 2020-09-03 2024-02-13 Lg Innotek Co., Ltd. Light path control member and display device comprising same

Citations (1)

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Publication number Priority date Publication date Assignee Title
US20060057556A1 (en) * 2002-10-21 2006-03-16 The Government Of The United States Of America Department Of Health And Human Services Contiguous capillary electrospray sources and analytical devices

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US20060057556A1 (en) * 2002-10-21 2006-03-16 The Government Of The United States Of America Department Of Health And Human Services Contiguous capillary electrospray sources and analytical devices

Non-Patent Citations (2)

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See also references of WO2012041767A1 *

Also Published As

Publication number Publication date
KR101516098B1 (ko) 2015-05-04
KR20130110173A (ko) 2013-10-08
JP2013537973A (ja) 2013-10-07
WO2012041767A1 (fr) 2012-04-05
JP5685651B2 (ja) 2015-03-18
CA2811109A1 (fr) 2012-04-05
DE102010041433A1 (de) 2012-03-29
US9410925B2 (en) 2016-08-09
US20140034496A1 (en) 2014-02-06

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