WO1999060394A1 - Colonne de rectification miniaturisee - Google Patents

Colonne de rectification miniaturisee Download PDF

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Publication number
WO1999060394A1
WO1999060394A1 PCT/EP1999/003086 EP9903086W WO9960394A1 WO 1999060394 A1 WO1999060394 A1 WO 1999060394A1 EP 9903086 W EP9903086 W EP 9903086W WO 9960394 A1 WO9960394 A1 WO 9960394A1
Authority
WO
WIPO (PCT)
Prior art keywords
separation
column
separating
miniaturized
capillary
Prior art date
Application number
PCT/EP1999/003086
Other languages
German (de)
English (en)
Inventor
Egbert Müller
Jürgen REGENER
Rudi Wollbeck
Original Assignee
Merck Patent Gmbh
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 Merck Patent Gmbh filed Critical Merck Patent Gmbh
Priority to JP2000549955A priority Critical patent/JP2002515604A/ja
Priority to EP99923534A priority patent/EP1078254A1/fr
Publication of WO1999060394A1 publication Critical patent/WO1999060394A1/fr

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/60—Construction of the column
    • G01N30/6052—Construction of the column body
    • G01N30/6073—Construction of the column body in open tubular form
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/14—Extraction; Separation; Purification
    • C07K1/16—Extraction; Separation; Purification by chromatography
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/50—Conditioning of the sorbent material or stationary liquid
    • G01N30/52—Physical parameters
    • G01N2030/524—Physical parameters structural properties
    • G01N2030/527—Physical parameters structural properties sorbent material in form of a membrane
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/60—Construction of the column
    • G01N30/6004—Construction of the column end pieces
    • G01N2030/6013—Construction of the column end pieces interfaces to detectors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/60—Construction of the column
    • G01N30/6004—Construction of the column end pieces
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/74—Optical detectors

Definitions

  • the invention relates to a miniaturized separation column, in particular for liquid chromatography, and to the use thereof for substance separation and in separation devices
  • Japanese Laid-Open Patent Application JP 63-246 657 discloses liquid-tightly coated flexible or rigid capillaries made of porous membrane materials which are suitable as miniaturized separation columns for gel permeation chromatography. References to the introduction of separation effectors are not given. This publication discloses separation - Sauces, in which sample and eluent contain the lumen of the hollow
  • miniaturized separating columns whose separating agents contain separation effectors and which are therefore suitable, for example, for ion exchange or reversed-phase chromatography.
  • These miniaturized separating columns are intended for tuned separation devices are suitable, can be produced reproducibly and are characterized by a low pressure drop
  • the invention relates to miniaturized chromatographic separation columns, the separation means of which are formed by a single porous hollow fiber membrane de ⁇ vatised with separation effectors, and which are provided with connections for eluent inflow and outflow two substances, and the use of the separation columns according to the invention in separation devices
  • FIGS 1, 2 and 3 (partial illustrations a and b) show
  • release agents are known which can also be formed as hollow fiber membranes. It has been found that individual porous hollow fibers can be used as release agents. Instead of a sorbent, such hollow fibers can be introduced are reproducibly producible. Chromatographic sauces provided with this separating agent allow separations in a wide range of linear flow velocities without an excessive pressure drop occurring
  • the term “separating agent” encompasses known particulate and non-particulate sorbents, and the separating membranes according to the invention, separating agents consist of a base support and separation effectors bound to this base support. Separation effectors effect the chromatographic separation of the substances in the eluent stream.
  • separation effectors are ionic groups such as the carboxyl or the sulfonic acid group for cation exchange chromatography, alkylated amino or ammonium groups for anion exchange chromatography, long and medium chain alkyl groups or aryl groups for reversed phase chromatography. Further examples of separation effectors are disclosed in WO 96/22 316 and WO 97/49 754 .
  • the separating membranes according to the invention are individual hollow fiber membranes, the separation effectors containing separating columns comprise, in addition to the separating means, a columnar column and
  • Connections for eluent inlet and outlet separation devices include, in addition to a separation column, an optional additional separation column as a pre-column, as well as pe ⁇ phere devices, which include, for example, eluent containers and pumps, gradient mixers, sample applicators and detectors
  • Hollow fiber membranes are generally manufactured with an outer diameter of 0.2 to 3 mm and an inner diameter of 0.1 to 1 mm.
  • Separation membranes are suitable for use in commercially available porous hollow fiber membranes, into which separation effectors have been introduced by suitable de ⁇ vatization methods.
  • the above-mentioned documents also disclose methods for de ⁇ vatization of polyamide membranes, according to which the said separation effectors can be introduced into the base membrane.
  • a separation membrane (hollow fiber membrane) is closed on one side, for example with an epoxy plug, and a capillary, for example made of polypropylene (PP), polyethylene (PE), polytetrafluoroethylene (PTFE), is inserted into the other end of the separation membrane. , Glass or also stainless steel inserted
  • the further structure of the separation column can be seen in Figures 1 to 3.
  • Figure 1 shows an embodiment of a separation column according to the invention
  • (1) denotes the column material, i.e. the porous hollow fiber, which is de ⁇ vated by separation effectors
  • (2) denotes that Casing tube
  • (3a) the capillary onto which the column material (1) is pushed
  • Capillary (3a) and pillar material (1) are fixed in a liquid-tight manner in the jacket tube (2) by means of a sleeve made of adhesive (4).
  • a plug made of adhesive (6) closes the lumen of Saulenmate ⁇ als, ie the porous hollow fiber (1)
  • Abla A capillary (3b) is provided, which is held in a sleeve made of adhesive (4) in the casing tube (2)
  • the capillary (3a) with the column material (1) and the capillary (3b) are not fixed with sleeves (4) made of glue, but with press rings made of steel (5)
  • Adhesive, or the pressing rings (5) made of steel, are replaced by screw connections ((7a) and 7b)) which have a double cone (8) made of deformable material around the capillary (3a) and the column material (1), as well as around the Press the capillary (3b)
  • the separation column thus comprises the separation membrane (1), ie a hollow fiber membrane closed on one side, the capillary right (3a) and the jacket tube (2), as well as sealing and connecting elements ((4), (5), (6), (7a), (7b), (8) and (3b))
  • the separation column according to the invention can also be integrated into a UV detector customary for HPLC (see Figure 4).
  • (9) means a screw with a central bore that connects the separation column
  • the embodiment according to Figure 4a has additional sealing elements on a PTFE cone (10) and a PTFE disk (11)
  • the dimensions of the jacket tube are selected so that the inner diameter of the jacket tube is slightly larger than the outside diameter of the separating membrane, so that only a small dead volume remains between the two.
  • the length of the jacket tube is chosen according to the length of the separating membrane
  • the separation column consists of electrically insulating materials, it can be used not only for chromatographic separations, but also for separations based on capillary electrophoresis or electrochromatography. Because of the low pressure drop during operation of the separation column according to the invention, it is also possible to use devices for others To provide separation methods that allow
  • capillary zone electrophoresis or isotachophoresis devices for connection and operation
  • additional separation processes are known in principle, for example from the coupling of chromatographic processes with the mass spectrometer.
  • Such devices can therefore be part of a separation device according to the invention as additional peripheral devices.
  • simple pumps are also possible How to use syringe pumps to demand the eluent
  • an additional separation column can also be installed as a preliminary column, as is usual in HPLC
  • Example 1 Preparation of a separation column for cation exchange chromatography (membrane of the SO 3 " type)
  • One end of a hollow fiber membrane made of polyamide modified according to WO 96/22 316 with SO 3 H groups is glued with Araldit "AW136H (Fa. Ciba-Geigy).
  • A136H Araldit
  • the separating membrane is inserted together with the steel capillary into a glass tube (inside diameter 0.6 mm), so that the steel capillary protrudes over the end of the glass tube.
  • a plug of Araldit ® AW136H (Fa. Ciba-Geigy) is produced, which both Steel capillary in the glass tube as well as the separating membrane attached to the steel capillary surrounds
  • a stainless steel capillary is fixed with a plug made of Araldit ® AW136H (from Ciba-Geigy)
  • the separation column manufactured according to Example 1 is installed in a separation device, the separation device comprises a gradient mixer for the two eluents, a sample application device and a UV detector.
  • the separation conditions were sample chymotrypsinogen, cytochrome C, lysozyme (15 ⁇ g / ⁇ l each) in 20 mM sodium phosphate buffer (pH 6 0) sample volume 5 ⁇ l Gradient:

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Physics & Mathematics (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Separation Using Semi-Permeable Membranes (AREA)

Abstract

L'invention concerne des colonnes de rectification miniaturisées dont le moyen de rectification est formé d'une membrane individuelle en fibre creuse poreuse ayant subi une dérivatisation avec des effecteurs de séparation, et qui sont munies de raccordements pour l'arrivée et l'évacuation d'agents d'élution.
PCT/EP1999/003086 1998-05-15 1999-05-05 Colonne de rectification miniaturisee WO1999060394A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2000549955A JP2002515604A (ja) 1998-05-15 1999-05-05 小型化分離カラム
EP99923534A EP1078254A1 (fr) 1998-05-15 1999-05-05 Colonne de rectification miniaturisee

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE19821812.5 1998-05-15
DE19821812A DE19821812C2 (de) 1998-05-15 1998-05-15 Miniaturisierte Trennsäule

Publications (1)

Publication Number Publication Date
WO1999060394A1 true WO1999060394A1 (fr) 1999-11-25

Family

ID=7867865

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1999/003086 WO1999060394A1 (fr) 1998-05-15 1999-05-05 Colonne de rectification miniaturisee

Country Status (4)

Country Link
EP (1) EP1078254A1 (fr)
JP (1) JP2002515604A (fr)
DE (1) DE19821812C2 (fr)
WO (1) WO1999060394A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002068452A2 (fr) * 2001-02-22 2002-09-06 Plant Bioscience Limited Procede d'isolement de composes charges

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE102005033425A1 (de) * 2005-07-18 2007-01-25 Omx Gmbh Verfahren zur Entfärbung von Proteinen
GB2538080A (en) 2015-05-05 2016-11-09 Endet Ltd Sorbent tube apparatus

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS61233367A (ja) * 1985-04-08 1986-10-17 Jeol Ltd クロマトグラフイ用マイクロカラムの製造方法
EP0369769A2 (fr) * 1988-11-15 1990-05-23 Hoechst Celanese Corporation Chromatographie liquide utilisant des fibres creuses microporeuses
WO1997049754A1 (fr) * 1996-06-21 1997-12-31 Merck Patent Gmbh Derives de polyamides polymerisables
DE19629208A1 (de) * 1995-01-20 1998-01-22 Merck Patent Gmbh Verwendung von modifizierten Membranen für "Simulated Moving Bed" Trennverfahren

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63246657A (ja) * 1987-03-31 1988-10-13 Chuichi Hirayama ゲルクロマトグラフイ−カラム
DE19501726A1 (de) * 1995-01-20 1996-07-25 Merck Patent Gmbh Polymerisationsfähige Derivate von Polyamiden

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS61233367A (ja) * 1985-04-08 1986-10-17 Jeol Ltd クロマトグラフイ用マイクロカラムの製造方法
EP0369769A2 (fr) * 1988-11-15 1990-05-23 Hoechst Celanese Corporation Chromatographie liquide utilisant des fibres creuses microporeuses
DE19629208A1 (de) * 1995-01-20 1998-01-22 Merck Patent Gmbh Verwendung von modifizierten Membranen für "Simulated Moving Bed" Trennverfahren
WO1997049754A1 (fr) * 1996-06-21 1997-12-31 Merck Patent Gmbh Derives de polyamides polymerisables

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
PATENT ABSTRACTS OF JAPAN vol. 11, no. 072 5 March 1987 (1987-03-05) *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002068452A2 (fr) * 2001-02-22 2002-09-06 Plant Bioscience Limited Procede d'isolement de composes charges
WO2002068452A3 (fr) * 2001-02-22 2003-10-16 Plant Bioscience Ltd Procede d'isolement de composes charges

Also Published As

Publication number Publication date
DE19821812C2 (de) 2000-03-16
DE19821812A1 (de) 1999-12-02
EP1078254A1 (fr) 2001-02-28
JP2002515604A (ja) 2002-05-28

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