US3869365A - Method in counter flow isotachophoresis - Google Patents

Method in counter flow isotachophoresis Download PDF

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Publication number
US3869365A
US3869365A US423997A US42399773A US3869365A US 3869365 A US3869365 A US 3869365A US 423997 A US423997 A US 423997A US 42399773 A US42399773 A US 42399773A US 3869365 A US3869365 A US 3869365A
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sample
column
counter flow
detector
electrolyte
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US423997A
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Bengt Fritiof Sunden
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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

Definitions

  • a method for fixing the sample in a certain pos tion in v 7 a counter flow isotachophoresis column comprises the 7 adjustment of voltage and counterflow values. to a 204/180 constant at which'the power from the electric field [58] Fie'ld G S and the power from the counterflow compensate each 1 204/299 other at the desired position of the sample.
  • PATENTEB AR 41975 sum 1 95 g ionized sample comprising ions of a certain polarity takes place by introducing the sample in a column arranged between two electrodes, a leading electrolyte, comprising ions of the same polarity but having a higher mobility than the sample, being introduced in the column between the sample and the electrodetowards which said ions migrate when a voltage is applied to the electrodes, and a terminating electrolyte comprising ions of said polarity having a lower mobility than that of the sample ions being introduced between the sample and the other electrode, the leading electrolyte being supplied to the column under pressure so as to bring the electrolyte to flow in a direction towards the sample.
  • Isotachophoresis is described more in detail, e.g. in Analytica Chemica Acta 38 (1967) pages 233-237 under the name ofDisplacement electrophoresis and is also described in the Swedish Pat. No.
  • 3,705,845 it is, furthermore, known to auto-' mate the adjustment procedure by using a detector which detects said boundary, and when the boundary moves compensates this movement by changing the amplitude of the counter flow.
  • the drawback of this method is that it requires an extra detector and electronic circuitry for controlling the counter flow, which means that the apparatus will be quite expensive.
  • the boundary is sharply defined since'this boundary is the parameter from which the regulation is based. Since counter flow isotachophoresis is mainly used when the components of the sample are difficult to separate this boundary will, at least in the beginning, be rather diffuse which makes the control uncertain.
  • FIG. 1 schematically shows the process of ion separa tion in isotachophoresis
  • FIG. 2 shows an apparatus for carrying 'out the method according to the invention
  • FIG. 3 by means of diagram explains the working principle of the apparatus according to FIG. 2.
  • reference 1 denotes a column in which an anode 5 and a cathode 4 are arranged. It is, furthermore, presumed that the sample to be separated is introduced in the part of the column denoted S, the sam- 'ple comprising two different anions C and C of which C is supposed to have a higher mobility than C
  • the part of the column denoted L is filled with the above described leading electrolytewhich consists of anions A having a higher mobility than all anions in the sample.
  • the part of the column T closed to the cathode is filled with an electrolyte comprising an anion B having a mobility which is lower than that of the anions in the sample.
  • a direct voltage is sup plied to the electrodes 4 and 5
  • the anions will migrate towards the anode 5. Because of the different mobility of the anions a zonewise and stepwise growing voltage gradient will be obtained across the zones L, S, and T,
  • the voltage gradient across the zone S will, however, imply that the ions within this zone are separated according to their mobility so that the ions C which have the higher mobility, are located close to the leading electrolyte and the ions C; with the lower mobility are located close to the terminating electrolyte.
  • the anions of the sample When a voltage is supplied to the column, the anions of the sample will thus be separated and after the separation, the different zones of the column will migrate towards the anode 5 with a velocity which is dependent upon the mobility of the ion A, a zonewise, growing potential being obtained across the column.
  • the thus formed zones will be very stable, since if an anion-from one zone e.g.
  • the anion will due to the lower potential gradient in the zone in front obtain a reduced velocity and be brought back into its original zone.
  • an anion which diffuses into a zone behind its original zone will be brought back to its original zone because of the higher voltage gradient in the zone behind.
  • the length of the column could, however, be considerably reduced if, during the separation, leading electrolyte is supplied to the column as a counter flow.
  • the amplitude of the counter flow could then preferably be chosen so as to keep the boundary between the zones L and S in a fixed position.
  • the control of the counter flow is either carried out manually by means of observations of the zone boundary and by increasing or reducing the counter'flow pressure when this boundary requirements of any controlling detector.
  • FIG. 2 there is shown schematically an apparatus for carrying outcounter flow isotachophoresis according to the invention.
  • reference 1 denotes a separation column in which the sample can be introduced via an input port 3 between a terminating and a leading electrolyte T and L, respectively.
  • A. voltage is applied to the column and the electrolyte by means of a voltage supply 2 which is connected to electrodes 4 and 5, respectively, whereby the sample S migrates into the column.
  • the power supply 2 is designed in such a column. Provided that a constant voltage V0 is applied to the column, the current will then successively decrease as the contents of leading electrolyte of the column gradually decreases whereas the contents of terminating electrolyte, i.e.
  • the column is further provided with a first detector 10 for detection of the separated zones. This detector is connected to a plotter 12 via an amplifier 11.
  • the apparatus is, fur thermore, provided with a second detector 7 which could be used for stabilizing the locations of the zones. This detector is in a corresponding manner connected to a plotter 9 via an amplifier 8.
  • FIG. 3a shows the current through a column accordingto FIG. 2 when a constant current V0 is applied to the electrodes as a function of time, and furthermore, the position of the sample Sin the column 1 during the process is indicated.
  • the essential principle of the invention is thus that by applying a column constant counter flow and a constant current to the column it is possible to fix the sample at an equilibrium where the current through the column is constant.
  • a current differential detector or some other conventional detector located along the column According to the invention one will thus obtain a process where it is very simple and unexpensive to fix the sample in a predetermined position during an arbitrary time.
  • the indication means consists of a thermodetector. 5. Method according to claim 3, characterized in, that the indication means consists of a UV-detector.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biochemistry (AREA)
  • Electrochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Investigating Or Analyzing Materials By The Use Of Electric Means (AREA)
  • Measuring Fluid Pressure (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
US423997A 1972-12-19 1973-12-12 Method in counter flow isotachophoresis Expired - Lifetime US3869365A (en)

Applications Claiming Priority (1)

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SE7216594A SE390766B (sv) 1972-12-19 1972-12-19 Forfarande vid motflodesisotachofores

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US (1) US3869365A (xx)
JP (1) JPS5653694B2 (xx)
DE (1) DE2363195A1 (xx)
FR (1) FR2226072A5 (xx)
GB (1) GB1454553A (xx)
SE (1) SE390766B (xx)

Cited By (25)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3941678A (en) * 1974-02-28 1976-03-02 Shimadzu Seisakusho Ltd. Apparatus for electrophoretic analysis
US3948753A (en) * 1973-11-13 1976-04-06 Lkb-Produkter Ab Apparatus for isotachophoretical separation
US3998719A (en) * 1974-08-21 1976-12-21 Ceskoslovenska Akademie Ved Isotachophoretic columns
US4459198A (en) * 1981-07-27 1984-07-10 Shimadzu Corporation Electrophoretic apparatus
US4617104A (en) * 1982-12-29 1986-10-14 Kureha Kagaku Kogyo Kabushiki Kaisha Cell unit for observing electrophoresis
US4666578A (en) * 1985-02-27 1987-05-19 Olympus Optical Co., Ltd. Method of measuring total protein of sample with the aid of electrophoretic image
US4666577A (en) * 1985-02-07 1987-05-19 Olympus Optical Co., Ltd. Method of recording electrophoretic image pattern
WO1989004966A1 (en) * 1987-11-25 1989-06-01 Norberto Guzman Automated capillary electrophoresis apparatus
EP0486559A1 (en) * 1989-08-07 1992-05-27 Applied Biosystems FRACTIONATION OF NUCLEIC ACID BY CAPILLARY ELECTROPHORESIS AGAINST MIGRATION.
US5275706A (en) * 1991-11-29 1994-01-04 Gerhard Weber Method and apparatus for continuous, carrier-free deflection electrophoresis
EP0608120A2 (en) * 1993-01-19 1994-07-27 Hewlett-Packard Company Capillary electrophoresis flow control system
US5429728A (en) * 1992-08-31 1995-07-04 Hewlett-Packard Company Electroosmotic flow control using back pressure in capillary electrophoresis
WO1998050787A1 (en) * 1997-05-08 1998-11-12 Sarnoff Corporation Indirect electrode-based pumps
US20100155241A1 (en) * 2006-10-04 2010-06-24 Ross David J Gradient elution electrophoresis
US20100224494A1 (en) * 2009-03-03 2010-09-09 The Board Of Trustees Of The Leland Stanford Junior University Isotachophoretic Focusing of Nucleic Acids
US20100261612A1 (en) * 2007-12-14 2010-10-14 Young Charles C Purification and Concentration of Proteins and DNA from a Complex Sample Using Isotachophoresis and a Device to Perform the Purification
US20100323913A1 (en) * 2007-12-14 2010-12-23 Young Charles C Purification and Concentration of Proteins and DNA from a Complex Sample Using Isotachophoresis and a Device to Perform the Purification
US20110174624A1 (en) * 2006-08-29 2011-07-21 Becton, Dickinson And Company Methods and Apparatus for Carrier-Free Deflection Electrophoresis
US20110220499A1 (en) * 2010-03-12 2011-09-15 Chambers Robert D Non-focusing tracers for indirect detection in electrophoretic displacement techniques
US8524061B2 (en) 2010-11-29 2013-09-03 The Board Of Trustees Of The Leland Stanford Junior University On-chip hybridization coupled with ITP based purification for fast sequence specific identification
US8562804B2 (en) 2006-07-20 2013-10-22 The Board Of Trustees Of The Leland Stanford Junior University Fluorescent finger prints for indirect detection in isotachophoresis
WO2014030997A1 (en) * 2012-08-21 2014-02-27 Universiteit Leiden Apparatus and process for depletion zone isotachophoresis
US8986529B2 (en) 2010-09-13 2015-03-24 The Board Of Trustees Of The Leland Stanford Junior University Isotachophoresis having interacting anionic and cationic shock waves
US10415030B2 (en) 2016-01-29 2019-09-17 Purigen Biosystems, Inc. Isotachophoresis for purification of nucleic acids
US11041150B2 (en) 2017-08-02 2021-06-22 Purigen Biosystems, Inc. Systems, devices, and methods for isotachophoresis

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5716115Y2 (xx) * 1976-06-30 1982-04-05
JPS58174389U (ja) * 1982-05-18 1983-11-21 スズキ株式会社 車両用燃料タンク
JPS59125497U (ja) * 1983-02-14 1984-08-23 スズキ株式会社 車両用燃料タンクの燃料移動防止装置

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3453200A (en) * 1966-05-25 1969-07-01 Instrumentation Specialties Co Apparatus for density gradient electrophoresis
US3649499A (en) * 1968-03-27 1972-03-14 Rauno Erkki Virtanen Method for establishing the zones occurring in electrophoresis and for their quantitative determination
US3649498A (en) * 1965-10-06 1972-03-14 Victor Pretorius Detection in chromatography
US3705845A (en) * 1970-06-02 1972-12-12 Lkb Produkter Ab Method in counterflow isotachophoresis
US3712859A (en) * 1968-06-13 1973-01-23 Ortec Inc Process for particle separation

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3649498A (en) * 1965-10-06 1972-03-14 Victor Pretorius Detection in chromatography
US3453200A (en) * 1966-05-25 1969-07-01 Instrumentation Specialties Co Apparatus for density gradient electrophoresis
US3649499A (en) * 1968-03-27 1972-03-14 Rauno Erkki Virtanen Method for establishing the zones occurring in electrophoresis and for their quantitative determination
US3712859A (en) * 1968-06-13 1973-01-23 Ortec Inc Process for particle separation
US3705845A (en) * 1970-06-02 1972-12-12 Lkb Produkter Ab Method in counterflow isotachophoresis

Cited By (42)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3948753A (en) * 1973-11-13 1976-04-06 Lkb-Produkter Ab Apparatus for isotachophoretical separation
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
US4459198A (en) * 1981-07-27 1984-07-10 Shimadzu Corporation Electrophoretic apparatus
US4617104A (en) * 1982-12-29 1986-10-14 Kureha Kagaku Kogyo Kabushiki Kaisha Cell unit for observing electrophoresis
US4666577A (en) * 1985-02-07 1987-05-19 Olympus Optical Co., Ltd. Method of recording electrophoretic image pattern
US4666578A (en) * 1985-02-27 1987-05-19 Olympus Optical Co., Ltd. Method of measuring total protein of sample with the aid of electrophoretic image
WO1989004966A1 (en) * 1987-11-25 1989-06-01 Norberto Guzman Automated capillary electrophoresis apparatus
EP0486559A1 (en) * 1989-08-07 1992-05-27 Applied Biosystems FRACTIONATION OF NUCLEIC ACID BY CAPILLARY ELECTROPHORESIS AGAINST MIGRATION.
EP0486559A4 (en) * 1989-08-07 1993-03-17 Applied Biosystems, Inc. Nucleic acid fractionation by counter-migration capillary electrophoresis
US5275706A (en) * 1991-11-29 1994-01-04 Gerhard Weber Method and apparatus for continuous, carrier-free deflection electrophoresis
US5429728A (en) * 1992-08-31 1995-07-04 Hewlett-Packard Company Electroosmotic flow control using back pressure in capillary electrophoresis
EP0608120A2 (en) * 1993-01-19 1994-07-27 Hewlett-Packard Company Capillary electrophoresis flow control system
EP0608120A3 (en) * 1993-01-19 1995-04-19 Hewlett Packard Co Flow control device for capillary electrophoresis.
US5482608A (en) * 1993-01-19 1996-01-09 Hewlett Packard Company Capillary electrophoresis flow control system
WO1998050787A1 (en) * 1997-05-08 1998-11-12 Sarnoff Corporation Indirect electrode-based pumps
US5961800A (en) * 1997-05-08 1999-10-05 Sarnoff Corporation Indirect electrode-based pumps
US8562804B2 (en) 2006-07-20 2013-10-22 The Board Of Trustees Of The Leland Stanford Junior University Fluorescent finger prints for indirect detection in isotachophoresis
US8795494B2 (en) 2006-08-29 2014-08-05 Becton, Dickinson And Company Methods and apparatus for carrier-free deflection electrophoresis
US20110174624A1 (en) * 2006-08-29 2011-07-21 Becton, Dickinson And Company Methods and Apparatus for Carrier-Free Deflection Electrophoresis
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
US20100323913A1 (en) * 2007-12-14 2010-12-23 Young Charles C Purification and Concentration of Proteins and DNA from a Complex Sample Using Isotachophoresis and a Device to Perform the Purification
US8865401B2 (en) 2007-12-14 2014-10-21 The Johns Hopkins University Purification and concentration of proteins and DNA from a complex sample using isotachophoresis and a device to perform the purification
US9377438B2 (en) 2007-12-14 2016-06-28 The Johns Hokpins University Kit for co-purification and concentration of DNA and proteins using isotachophoresis
US20100261612A1 (en) * 2007-12-14 2010-10-14 Young Charles C Purification and Concentration of Proteins and DNA from a Complex Sample Using Isotachophoresis and a Device to Perform the Purification
US8614059B2 (en) 2007-12-14 2013-12-24 The Johns Hopkins University Purification and concentration of proteins and DNA from a complex sample using isotachophoresis and a device to perform the purification
US9753007B1 (en) 2009-03-03 2017-09-05 The Board Of Trustees Of The Leland Stanford Junior University Isotachophoretic focusing of nucleic acids
US20100224494A1 (en) * 2009-03-03 2010-09-09 The Board Of Trustees Of The Leland Stanford Junior University Isotachophoretic Focusing of Nucleic Acids
US8846314B2 (en) 2009-03-03 2014-09-30 The Board Of Trustees Of The Leland Stanford Junior University Isotachophoretic focusing of nucleic acids
US8721858B2 (en) 2010-03-12 2014-05-13 The Board Of Trustees Of The Leland Stanford Junior University Non-focusing tracers for indirect detection in electrophoretic displacement techniques
US20110220499A1 (en) * 2010-03-12 2011-09-15 Chambers Robert D Non-focusing tracers for indirect detection in electrophoretic displacement techniques
US8986529B2 (en) 2010-09-13 2015-03-24 The Board Of Trustees Of The Leland Stanford Junior University Isotachophoresis having interacting anionic and cationic shock waves
US8524061B2 (en) 2010-11-29 2013-09-03 The Board Of Trustees Of The Leland Stanford Junior University On-chip hybridization coupled with ITP based purification for fast sequence specific identification
US9575031B2 (en) 2012-08-21 2017-02-21 Universiteit Leiden Apparatus and process for depletion zone isotachophoresis
WO2014030997A1 (en) * 2012-08-21 2014-02-27 Universiteit Leiden Apparatus and process for depletion zone isotachophoresis
US10415030B2 (en) 2016-01-29 2019-09-17 Purigen Biosystems, Inc. Isotachophoresis for purification of nucleic acids
US10822603B2 (en) 2016-01-29 2020-11-03 Purigen Biosystems, Inc. Isotachophoresis for purification of nucleic acids
US11674132B2 (en) 2016-01-29 2023-06-13 Purigen Biosystems, Inc. Isotachophoresis for purification of nucleic acids
US12006496B2 (en) 2016-01-29 2024-06-11 Purigen Biosystems, Inc. Isotachophoresis for purification of nucleic acids
US11041150B2 (en) 2017-08-02 2021-06-22 Purigen Biosystems, Inc. Systems, devices, and methods for isotachophoresis
US11987789B2 (en) 2017-08-02 2024-05-21 Purigen Biosystems, Inc. Systems, devices, and methods for isotachophoresis

Also Published As

Publication number Publication date
JPS4991495A (xx) 1974-08-31
DE2363195A1 (de) 1974-06-27
FR2226072A5 (xx) 1974-11-08
SE390766B (sv) 1977-01-17
DE2363195B2 (xx) 1975-07-03
JPS5653694B2 (xx) 1981-12-21
GB1454553A (en) 1976-11-03

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