US3705845A - Method in counterflow isotachophoresis - Google Patents

Method in counterflow isotachophoresis Download PDF

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Publication number
US3705845A
US3705845A US148657A US3705845DA US3705845A US 3705845 A US3705845 A US 3705845A US 148657 A US148657 A US 148657A US 3705845D A US3705845D A US 3705845DA US 3705845 A US3705845 A US 3705845A
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sample
ions
column
electrolyte
counterflow
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US148657A
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Frans Everaerts
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Pfizer Health AB
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LKB Produkter AB
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    • 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
    • G01N27/44765—Apparatus specially adapted therefor of the counter-flow type

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  • FIG. 1a A first figure.
  • FIG. 2 RIG 1' F. EVERAERTS METHOD IN COUNTERFLOW ISOTACHOPHORESIS Filed June 1, 1971 DecQ 12, 1972 FIG. 2
  • first and second electrolytes comprise ions of higher and lower mobility respectively than the ions of said sample.
  • a voltage is applied to the electrodes and a pressure is applied to one of said electrolytes.
  • a boundary between zones of ions having different mobilities is detected and from this detection a control signal is generated, said control signal controlling said pressure and/or said voltage.
  • the present invention relates to a method in counterflow isotachophoresis, in which a sample, containing ions of the same polarity which are to be separated, is introduced into a column arranged between two electrodes, at which in that part of the column which is situated between the sample and that electrode towards which the ions in question are migrating when a voltage is applied to the electrodes, a leading electrolyte is introduced containing ions of still the same polarity but with a higher mobility than that of the sample ions, and between the sample and the other electrode a terminating electrolyte is introduced containing ions of still the same polarity but with a lower mobility than that of the sample ions, and that the above mentioned leading electrolyte is supplied to the column under a pressure so that the electrolyte is made flowing towards the sample.
  • Isotachophoresis is principally carried out in that way that a sample containing a number of different ions with the same polarity is introduced into some type of column in which the sample is located between two electrolytes, containing ions of the same polarity as those ions in the sample which are going to be separated, whereat one of the electrolytes contains ions with a higher mobility than that of the sample ions and the other electrolyte contains ions with a lower mobility than that of the sample ions.
  • the electrolyte containing ions with a higher mobility is at that introduced in that part of the column which is connected to the electrode, towards which the sample ions are migrating, and the electrolyte with the slower ions, the so called terminating electrolyte, is introduced at the other electrode. If a voltage is then applied to the electrodes, the ions will be migrating in the column, at which the ions of the sample will be separated according to their mobility. The course thus obtained will be closer explained below.
  • the essential advantage of isotachophoresis in comparison with conventional electrophoresis is that substantially smaller amounts of sample can be separated and that very sharp boundaries are achieved between the separated zones of sample, the zones of sample thus not be- Patented Dec. 12, 1972 ing extended during the course of separation as in conventional electrophoresis.
  • a disadvantage is, however, that a long column is to be used in working with very small ions concentrations and in working with sample ions with very slight diiference in mobility, whereby very high potentials are required to give sufficient field strength in the column.
  • the length of the column might, however, be considerably reduced if a counterflow of the leading electrolyte is utilized, i.e. the leading electrolyte is pumped against the migration direction of the sample ions. (See e.g.
  • the purpose of the present invention is to provide a method in counterflow isotachophoresis in which those disadvantages are eliminated.
  • FIGS. la and 1b schematically show how the different ions of a sample are separated
  • FIG. 2 also schematically, shows a device for accomplishing the method according to the invention.
  • the number 1 denotes a column in which an anode 5 and a cathode 4 are introduced. It is further assumed that the sample which is to be separated is introduced into that part of the column which is designated with S, and that the sample consists of salts containing two diiferent anions C and C of which C is assumed to have a higher mobility than C and a common cation R+.
  • the part of the column which is designated L is filled with the above mentioned leading electrolyte, which consists of anions A with a higher mobility than all the anions of the sample and a cation, designated R+, suitably common with the sample.
  • the part T of the column which is turned towards the cathode is filled with an electrolyte which contains an anion B-, the mobility of which is lower than of all anions of the sample, and a cation R+, common with the leading electrolyte.
  • an electrolyte which contains an anion B-, the mobility of which is lower than of all anions of the sample, and a cation R+, common with the leading electrolyte.
  • the correspondingly stepwise increasing temperature on the outer surface of the column 5 could be measured or the potential at one or more points on the column could be directly measured.
  • the electrode it is also possible, by means of the electrode, to measure the conductivity of the zones passing a certain point, and so far the separated substances are UV-absorbing, the UV- absorption could also be measured in conventional manner.
  • the length of the column can, however, be considerably reduced, if leading electrolyte is continuously pumped to the column during the separation.
  • the magnitude of the counterflow can then suitably be chosen in a way that the boundary between the zones L and S principally is taining leading electrolyte has been raised or lowered.
  • the purpose of the present invention is therefore to provide a procedure by means of which the counterflow always is given such a value that a certain zone boundary is occupying a fixed position.
  • the number 1 denotes a column, which is arranged between an anode chamber 3 and a cathode chamber 2.
  • an anode 5 and a cathode 4 are respectively introduced and these are connected to a current source 6.
  • the column is further equipped with a multiway valve 7 by means of which the column could either be connected to the chamber 2 or to a sample injecting means 8.
  • the injecting means 30 the sample can be introduced into a capillary between the terminating electrolyte, which is introduced into the cathode chamber 2, and the leading electrolyte, which is introduced into the anode chamber 3 and in addition into that part of the column which is situated between the 5 valve and this chamber.
  • That part of the column which is situated at the anode is further suitably equipped with a valve 10 via which leading electrolyte could be supplied or removed. Further, a diaphragm 20 is arranged between the column and the anode chamber 5 in a way that the column could be flushed with liquid without changing the circumstances in the chamber. With those polarities of the electrodes 4 and 5 which have been chosen in the figure, it is then assumed that the anions of the sample are the subjects of separation. If the sample on the other hand would contain a number of diflerent cations the reverse polarity of the electrodes would of course be chosen. The device is further equipped with means for varying the counterflow of the leading electrolyte.
  • This means consist of a reservoir 11, in which a floater 13 can be moved up and down so that the liquid level in the reservoir and thus the pressure of the counterflow can be varied.
  • a spindle 14 which is supporting the floater is introduced into an electromagnet 15 so that by varying the current in the electromagnet the floater is brought up and down in the reservoir.
  • the governing of the current through the electromagnet is accomplished by means of a detection means 17 connected to the column.
  • the detection organ can be thermocouple but can also consist of any other of the detecting elements which has been stated above in connection to FIGS. la and lb.
  • the thermocouple is connected to an amplifier 18, the output of which in its turn is connected to a governing device 19. This one is in its turn connected to the electromagnet 15.
  • the signal from the thermocouple should then govern the counterflow in order that a zone boundary, suitably the boundary between'the leading electrolyte and the first sample zone, is mainly stationary at the thermocouple. As hinted above there will be obtained a temperature rise at this boundary which temperature rise also will cause a temperature rise on the outer surface of the column.
  • the governing device 19 should principally give an output signal which keeps the floater 13 in a fixed position when the signal from the thermocouple corresponds to a temperature which constitutes the mean value between the temperature in the leading electrolyte and that in the first sample zone.
  • a signal corresponding to a lower temperature should further cause a slow decreasing of the coun terflow by means of raising the floater 13, and a signal corresponding to a higher temperature should correspondingly cause a slow increase of the counterflow, i.e. a lowering of the floater 13.
  • counterflow governing by means of a vertically adjustable floater of course only is one example of varying the eounterflow.
  • a great advantage of the shown counterflow governing is, however, that very small counterflow alterations, free from pulsation, can be achieved, to a very inconsiderable extent disturbing the separation in the column.
  • any pressure generating means could be generally utilized.
  • the governing device 19 is also connected to a current source 6, and the purpose of this connection is that by varying the electric current in the column in dependence of the current from the thermocouple a certain zone boundary should be kept in a fixed position. Of course it is also possible to fix the zone boundary by alfecting both the counterflow and the electric current.
  • Method in counterflow isotachophoresis comprising the steps of:

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
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  • Electrochemistry (AREA)
  • Physics & Mathematics (AREA)
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US148657A 1970-06-02 1971-06-01 Method in counterflow isotachophoresis Expired - Lifetime US3705845A (en)

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SE07624/70A SE340376B (fr) 1970-06-02 1970-06-02
US14865771A 1971-06-01 1971-06-01

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Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3869365A (en) * 1972-12-19 1975-03-04 Lkb Produkter Ab Method in counter flow isotachophoresis
US3912609A (en) * 1973-01-15 1975-10-14 Lkb Produkter Ab Method at isotachophoretical separation to detect spectrophotometrically zone boundaries obtained
US3915827A (en) * 1973-01-24 1975-10-28 Lkb Produkter Ab Method in electrophoresis for controlling the supplied power so as to obtain separations at an optimum speed
US3941678A (en) * 1974-02-28 1976-03-02 Shimadzu Seisakusho Ltd. Apparatus for electrophoretic analysis
US3998719A (en) * 1974-08-21 1976-12-21 Ceskoslovenska Akademie Ved Isotachophoretic columns
US4290855A (en) * 1979-12-31 1981-09-22 The Regents Of The University Of California Method of isotope enrichment
US4323439A (en) * 1979-12-31 1982-04-06 The Regents Of The University Of California Method and apparatus for dynamic equilibrium electrophoresis
US4617104A (en) * 1982-12-29 1986-10-14 Kureha Kagaku Kogyo Kabushiki Kaisha Cell unit for observing electrophoresis
US5234559A (en) * 1991-12-31 1993-08-10 E. I. Du Pont De Nemours And Company Apparatus for direct blotting and automated electrophoresis, transfer and detection and processes utilizing the apparatus thereof
US5302264A (en) * 1992-09-02 1994-04-12 Scientronix, Inc. Capillary eletrophoresis method and apparatus
EP0608120A2 (fr) * 1993-01-19 1994-07-27 Hewlett-Packard Company Système de contrôle de débit en électrophorèse capillaire
US5429728A (en) * 1992-08-31 1995-07-04 Hewlett-Packard Company Electroosmotic flow control using back pressure in capillary electrophoresis
US5472584A (en) * 1991-10-04 1995-12-05 The Dow Chemical Company Method and apparatus for improved detection of ionic species by capillary electrophoresis
US20100155241A1 (en) * 2006-10-04 2010-06-24 Ross David J Gradient elution electrophoresis
FR3030305A1 (fr) * 2014-12-22 2016-06-24 Commissariat Energie Atomique Mesure isotopique multielementaire par couplage direct d'une technique d'isotachophorese multi-cycles et d'une technique de spectrometrie de masse.

Cited By (20)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3869365A (en) * 1972-12-19 1975-03-04 Lkb Produkter Ab Method in counter flow isotachophoresis
US3912609A (en) * 1973-01-15 1975-10-14 Lkb Produkter Ab Method at isotachophoretical separation to detect spectrophotometrically zone boundaries obtained
US3915827A (en) * 1973-01-24 1975-10-28 Lkb Produkter Ab Method in electrophoresis for controlling the supplied power so as to obtain separations at an optimum speed
US3941678A (en) * 1974-02-28 1976-03-02 Shimadzu Seisakusho Ltd. Apparatus for electrophoretic analysis
US3998719A (en) * 1974-08-21 1976-12-21 Ceskoslovenska Akademie Ved Isotachophoretic columns
US4290855A (en) * 1979-12-31 1981-09-22 The Regents Of The University Of California Method of isotope enrichment
US4323439A (en) * 1979-12-31 1982-04-06 The Regents Of The University Of California Method and apparatus for dynamic equilibrium electrophoresis
US4617104A (en) * 1982-12-29 1986-10-14 Kureha Kagaku Kogyo Kabushiki Kaisha Cell unit for observing electrophoresis
US5472584A (en) * 1991-10-04 1995-12-05 The Dow Chemical Company Method and apparatus for improved detection of ionic species by capillary electrophoresis
US5234559A (en) * 1991-12-31 1993-08-10 E. I. Du Pont De Nemours And Company Apparatus for direct blotting and automated electrophoresis, transfer and detection and processes utilizing the apparatus thereof
US5429728A (en) * 1992-08-31 1995-07-04 Hewlett-Packard Company Electroosmotic flow control using back pressure in capillary electrophoresis
US5302264A (en) * 1992-09-02 1994-04-12 Scientronix, Inc. Capillary eletrophoresis method and apparatus
EP0608120A2 (fr) * 1993-01-19 1994-07-27 Hewlett-Packard Company Système de contrôle de débit en électrophorèse capillaire
EP0608120A3 (fr) * 1993-01-19 1995-04-19 Hewlett Packard Co Système de contrÔle de débit en électrophorèse capillaire.
US5482608A (en) * 1993-01-19 1996-01-09 Hewlett Packard Company Capillary electrophoresis flow control system
US20100155241A1 (en) * 2006-10-04 2010-06-24 Ross David J Gradient elution electrophoresis
US8080144B2 (en) 2006-10-04 2011-12-20 The United States of America as represented by the Secretary of Commerce, the National Institute of Standards and Technology Gradient elution electrophoresis
FR3030305A1 (fr) * 2014-12-22 2016-06-24 Commissariat Energie Atomique Mesure isotopique multielementaire par couplage direct d'une technique d'isotachophorese multi-cycles et d'une technique de spectrometrie de masse.
EP3037153A1 (fr) * 2014-12-22 2016-06-29 Commissariat A L'energie Atomique Et Aux Energies Alternatives Mesure isotopique multielementaire par couplage direct d'une technique d'isotachophorese multi-cycles et d'une technique de spectrometrie de masse
US9437411B2 (en) 2014-12-22 2016-09-06 Commissariat A L'energie Atomique Et Aux Energies Alternatives Multi-element isotopic measurement by direct coupling of a multi-cycle isotachophoresis technique and a mass spectrometry technique

Also Published As

Publication number Publication date
GB1333796A (en) 1973-10-17
DE2127391B2 (de) 1972-07-06
FR2093980A1 (fr) 1972-02-04
DE2127391A1 (de) 1971-12-23
FR2093980B1 (fr) 1975-07-04

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