EP2192963A1 - Assemblages de colonnes de chromatographie - Google Patents

Assemblages de colonnes de chromatographie

Info

Publication number
EP2192963A1
EP2192963A1 EP08795368A EP08795368A EP2192963A1 EP 2192963 A1 EP2192963 A1 EP 2192963A1 EP 08795368 A EP08795368 A EP 08795368A EP 08795368 A EP08795368 A EP 08795368A EP 2192963 A1 EP2192963 A1 EP 2192963A1
Authority
EP
European Patent Office
Prior art keywords
column
particle size
average particle
column assembly
packing
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Withdrawn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
EP08795368A
Other languages
German (de)
English (en)
Inventor
Scott C. Anderson
Laura T. Kaepplinger
Juergen T. Maier-Rosenkranz
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.)
All-Tech Associates Inc
Original Assignee
All-Tech Associates Inc
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 All-Tech Associates Inc filed Critical All-Tech Associates Inc
Publication of EP2192963A1 publication Critical patent/EP2192963A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
    • B01D15/08—Selective adsorption, e.g. chromatography
    • B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
    • B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
    • B01D15/1864—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns
    • 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
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/38—Flow patterns
    • G01N30/46—Flow patterns using more than one column
    • G01N30/461—Flow patterns using more than one column with serial coupling of separation columns
    • 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
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G01N30/08—Preparation using an enricher
    • G01N2030/085—Preparation using an enricher using absorbing precolumn

Definitions

  • the present invention is directed to chromatography column assemblies, methods of making chromatography column assemblies, and methods of using chromatography column assemblies.
  • Guard columns are used in combination with an analytical or separation column to remove impurities and contaminants from a test sample prior to the test sample entering the analytical or separation column. By removing impurities and contaminants from a test sample prior to the test sample entering the analytical column, the useful lifetime of the analytical column is extended, and the effectiveness of the analytical column is improved. [0003] Typically, in ultra high pressure liquid chromatography
  • the particle size of packing media used in the guard column is equal to the particle size of packing media used in the analytical column in order to maintain a desired level of system efficiency, typically measured by the number of theoretical plates within a given column assembly.
  • a column assembly results in increased back pressure and reduced flow rates within the column assembly.
  • the chromatography column assembly comprises (i) an analytical column comprising a first packing media having a first average particle size, and (ii) a guard column in-line with the analytical column, the guard column comprising a second packing media having a second average particle size, the second average Docket No. W9795-01
  • the chromatography column assembly having a system efficiency greater than a similar chromatography column assembly in which the second average particle size is equal to or less than the first average particle size.
  • the present invention is also directed to methods of making chromatography column assemblies.
  • the method of making a chromatography column assembly comprises combining (i) an analytical column comprising a first packing media having a first average particle size with (ii) a guard column so that the guard column is in-line with the analytical column, wherein the guard column comprises a second packing media having a second average particle size that is greater than the first average particle size such that the resulting chromatography column assembly has a system efficiency greater than a similar chromatography column assembly in which the second average particle size is equal to or less than the first average particle size.
  • the present invention is further directed to methods of improving a system efficiency of a chromatography column assembly.
  • the method of improving a system efficiency of a chromatography column assembly comprises providing an analytical column comprising a first packing media having a first average particle size; and connecting a guard column to the analytical column to form a chromatography column assembly, the guard column comprising a second packing media having a second average particle size, the second average particle size being greater than the first average particle size; wherein the chromatography column assembly has a system efficiency greater than a similar chromatography column assembly in which the second average particle size is equal to or less than the first average particle size
  • the present invention is even further directed to methods of using chromatography column assemblies.
  • the method comprises a method of analyzing a test sample that potentially contains at least one analyte, wherein the method comprises the step of introducing the test sample into a chromatography column assembly comprising (i) an analytical column comprising a first packing media Docket No. W9795-01
  • the chromatography column assembly has a system efficiency greater than a similar chromatography column assembly in which the second average particle size is equal to or less than the first average particle size.
  • FIG. 1 depicts an exemplary chromatography column assembly of the present invention.
  • the present invention is directed to chromatography column assemblies comprising a guard column in combination with an analytical column, wherein the packing media of the guard column has a greater average particle size than the packing media of the analytical column, and the chromatography column assembly has a system efficiency greater than a similar chromatography column assembly in which the packing media of the guard column has an average particle size equal to or less than the packing media of the analytical column.
  • the present invention is further directed to methods of making chromatography column assemblies, as well as methods of using Docket No. W9795-01
  • exemplary chromatography column assembly 10 comprises a guard column 11; packing media 12 within guard column 11; an analytical column 13 positioned in-line with guard column 11; and packing media 14 within analytical column 13.
  • guard column 11 is shown separate from analytical column 13, it should be understood that guard column 11 and analytical column 13 may be integrally joined to one another (e.g., guard column 11 and analytical column 13 may share a column wall).
  • the chromatography column assemblies of the present invention may comprise a number of components. A description of possible components and various configurations is provided below.
  • the chromatography column assemblies of the present invention comprise a number of components, which result in a chromatography column assembly having unexpectedly superior system efficiency.
  • the chromatography column assemblies of the present invention comprise one or more of the following components.
  • the chromatography column assemblies of the present invention comprise at least one analytical column such as analytical column 13 of exemplary chromatography column assembly 10 shown in FIG. 1.
  • analytical columns may be used in the present invention depending on a number of factors including, but not limited to, the chemistry of the sample to be tested, separation goals (e.g., to separate one or more component(s) from any remaining components), speed of analysis, a desired number of theoretical plates, a desired resolution of one or more components in the sample, column stability and expected lifetime, and column sensitivity.
  • Analytical columns suitable for use in the present invention may have a variety of sizes depending on the intended use of the chromatography column assembly.
  • analytical columns suitable for use in the present invention may have any height (also referred to herein as the column length), although analytical columns typically have an overall height of up to about 3 meters (m).
  • analytical columns used in the present invention have a height (or length) ranging from about 30 mm to about 1.0 m.
  • analytical columns used in the present invention have a height (or length) ranging from about 50 mm to about 300 mm.
  • Analytical columns operatively adapted for use in the present invention have a cross-sectional flow area that may vary in size depending on the intended use of a given column.
  • the cross-sectional flow area is up to about 6.0 mm 2 .
  • the cross-sectional flow area ranges from about 0.075 mm 2 to about 5.0 mm 2 .
  • the cross-sectional flow area ranges from about 2.0 mm to about 4.6 mm .
  • Analytical columns operatively adapted for use in the present invention may be constructed from a variety of materials (e.g., stainless steel, aluminum, fiber-reinforced composite materials, PEEK, PTFE, or combinations thereof) in order to withstand an internal pressure that varies depending on the end use of a given column.
  • materials e.g., stainless steel, aluminum, fiber-reinforced composite materials, PEEK, PTFE, or combinations thereof
  • analytical columns operatively adapted for use in the present invention are constructed to have a pressure capacity of up to about 1034 bar (15,000 psig).
  • analytical columns operatively adapted for use in the present invention are constructed to have a pressure capacity ranging from about 34 bar (500 psig) to about 345 bar (5,000 psig).
  • a number of analytical columns operatively adapted for use in the present invention are commercially available from a variety of sources. Suitable analytical columns for use in the present invention include, but are not limited to, PlatinumTM HPLC columns and VisionHTTM HPLC columns commercially available from Grace Davison Discovery Sciences (Columbia, MD).
  • the analytical column used in the chromatography column assembly of the present invention Docket No. W9795-01
  • Each analytical column comprises a separation or packing media suitable for use in a given analytical column such as exemplary packing media 12 shown in FIG. 1.
  • packing media may be used in the present invention depending on a number of factors including, but not limited to, the factors listed above for column selection, column bed dimensions, particle size, particle surface area, particle pore size, particle chemical composition, surface treatments of particles, and bonding type (e.g., monomeric versus polymeric).
  • Any separation or packing media used in analytical columns may be used in the present invention.
  • the packing media comprises inorganic substances commercially available as chromatographic media. These substances can be prepared using methods known in the art.
  • the inorganic substances used in the present invention comprise an inorganic oxide, more suitably an inorganic metal oxide, silicate or aluminosilicate.
  • Inorganic oxides suitable for the present invention have free hydroxyl groups capable of bonding to or reacting with other chemical functionalities.
  • desirable inorganic metal oxides include silica such as chromatographic grade silica or silica gel, alumina, silica-alumina, zirconia, zirconate, controlled pore glass or titania.
  • the inorganic metal oxide is silica, more desirably chromatographic grade silica or silica gel.
  • Magnetically responsive inorganic metal oxides such as siliceous oxide-coated magnetic particles disclosed in WO 98/31461, the disclosure of which is incorporated by reference, are also suitable.
  • Mixed inorganic metal oxides e.g. co-gels of silica and alumina, or co-precipitates can also be used.
  • Solids prepared from sodium silicate are examples of a suitable silicate and zeolite is an example of a suitable aluminosilicate.
  • the inorganic substances used in the present invention can be in a physical form of particulates, fibers and plates. [0026]
  • the inorganic substances used in the present invention may further comprise one or more surface moieties, binding moieties, and/or linkers bonded to an outer surface of the inorganic substances, Docket No. W9795-01
  • inorganic oxides include from about 1 to about 10 hydroxyl groups per square nanometer of solid inorganic oxide for use in bonding to surface moieties, binding moieties, and/or linkers.
  • Suitable types of separation or packing media include, but are not limited to, inorganic particles with or without one or more surface moieties, binding moieties, and/or linkers such as those disclosed in U.S. Patent No. 6,802,966, the disclosure of which is incorporated herein by reference in its entirety.
  • the average particle size of the above- mentioned particles can vary depending on a particular application, the average particle size of suitable packing media for use in analytical columns of the column assemblies of the present invention is typically less than about 10.0 ⁇ m (or less than about 9.0 ⁇ m, or less than about 8.0 ⁇ m, or less than about 7.0 ⁇ m, or less than about 6.0 ⁇ m, or less than about 5.0 ⁇ m, or less than about 4.0 ⁇ m) more typically, less than about 3 ⁇ m. In one desired embodiment, the average particle size of suitable packing media for use in analytical columns of the column assemblies of the present invention is from about 1.0 ⁇ m to about 2.5 ⁇ m.
  • the average particle size of suitable packing media for use in analytical columns of the column assemblies of the present invention is about 1.5 ⁇ m.
  • the chromatography column assembly comprises an analytical column containing rigid support media, wherein the rigid support media comprises a plurality of spherically-shaped silica-based inorganic particles having a particle surface area of about 200 m 2 /g, an average pore size of about 100 angstroms, and an average particle size of about 1.5 ⁇ m or 3.0 ⁇ m, more typically, about 1.5 ⁇ m.
  • the silica-based inorganic particles may be further modified by chemically bonding a surface treatment (e.g., octadecyl silane) to the porous surface of the silica-based inorganic particles.
  • a surface treatment e.g., octadecyl silane
  • suitable surface moieties, binding moieties, and/or linkers are disclosed, for example, in U.S. Patent No. 6,802,966. Docket No. W9795-01
  • the chromatography column assemblies of the present invention further comprise at least one guard column such as exemplary guard column 13 of exemplary chromatography column assembly 10 shown in FIG. 1.
  • a variety of guard columns may be used in the present invention depending on a number of factors including, but not limited to, the chemistry of the sample to be tested, potential contaminants and/or impurities in the sample to be tested, the analytical column used, the desired speed of analysis, the desired number of theoretical plates, the desired back pressure, and the desired flow rate.
  • Guard columns suitable for use in the present invention have dimensions, cross-sectional flow areas, and column constructions similar to those described above for analytical columns suitable for use in the present invention.
  • guard columns used in the present invention have a height (or length) ranging from about 5.0 mm to about 1.0 m, more typically, from about 5.0 to about 10 mm, and a cross-sectional flow area ranging from about 0.075 mm 2 to about 4.6 mm 2 .
  • guard columns operatively adapted for use in the present invention are soon to be commercially available from Grace Davison Discovery Sciences (Columbia, MD).
  • Suitable guard columns for use in the present invention include, but are not limited to, PlatinumTM HPLC columns and VisionHTTM HPLC columns soon to be commercially available from Grace Davison Discovery Sciences (Columbia, MD).
  • the guard column used in the chromatography column assembly of the present invention comprises a VisionHTTM HPLC column or a PlatinumTM HPLC column.
  • Each guard column comprises a separation or packing media suitable for use in a given guard column such as exemplary packing media 14 shown in FIG. 1.
  • a variety of packing media may be used in a given guard column depending on a number of factors including, but not limited to, the factors listed above for analytical Docket No. W9795-01
  • any separation or packing media typically used in guard columns may be used in the present invention. Suitable types of separation or packing media include, but are not limited to, the packing media described above for use in analytical columns.
  • the average particle size of the above- mentioned particles can vary depending on a particular application, the average particle size of suitable packing media for use in guard columns of the column assemblies of the present invention is typically at least about 3.0 ⁇ m (or at least about 3.5 ⁇ m, or at least about 4.0 ⁇ m, or at least about 4.5 ⁇ m, or at least about 5.0 ⁇ m, or at least about 5.5 ⁇ m, or at least about 6.0 ⁇ m) up to about 100 ⁇ m. In one desired embodiment, the average particle size of suitable packing media for use in guard columns of the column assemblies of the present invention is from about 3.0 ⁇ m to about 10 ⁇ m. In another desired embodiment, the average particle size of suitable packing media for use in guard columns of the column assemblies of the present invention is about 3.0 ⁇ m.
  • the chromatography column assembly comprises a guard column containing rigid support media, wherein the rigid support media comprises a plurality of spherically-shaped silica-based inorganic particles having a particle surface area of about 200 m 2 /g, an average pore size of about 100 angstroms, and an average particle size of about 3.0 ⁇ m.
  • the silica-based inorganic particles may be further modified by chemically bonding a surface treatment (e.g., octadecyl silane) to the porous surface of the silica-based inorganic particles.
  • the chromatography column assemblies of the present invention may further comprise a number of conventional components used in uHPLC applications. Additional conventional components include, but are not limited to, a sample injector device operatively adapted to enable injection of a sample into the chromatography column assembly; one or more pistons within a given column so as to Docket No. W9795-01
  • system efficiency of a given column or chromatography column assembly of the present invention is measured by determining a number of theoretical plates, N, within the column or column assembly.
  • the number of theoretical plates, N, for a given column or column assembly may be determined as follows:
  • t R 2 is retention time of a peak; and w 1/2 is a peak width at a half-height of the peak.
  • the number of theoretical plates, N, per meter (m) of column length, N/m may be obtained by dividing the number of theoretical plates, N, by the length of the column (or the total length of columns within a column assembly).
  • the number of theoretical plates, N, per meter (m) of column length for exemplary guard columns for use in chromatography column assemblies of the present invention and containing packing material with an average particle size of about 3 ⁇ m is at least 3% greater than a similar guard column in which the average particle size of packing media within the guard column is about 1.5 ⁇ m.
  • the number of theoretical plates, N, per meter (m) of column length for exemplary guard columns for use in chromatography column assemblies of the present invention and containing packing material with an average particle size of about 3 ⁇ m is at least 4% greater (or at least 5% greater, or at least 6% greater, or at least 7% greater, or at least 8% greater, or at least 9% greater, or at least 10% greater) than a similar guard column in which the average particle size of packing media within the guard column is about 1.5 ⁇ m.
  • the number of theoretical plates, N, per meter (m) of column length for exemplary chromatography column assemblies of the present invention is at least 1% greater than a similar chromatography column assembly in which the average particle size of packing media within the guard column is equal to or less than the average particle size of packing media within the analytical column.
  • the number of theoretical plates, N, per meter (m) of column length, Mm, for exemplary chromatography column assemblies of the present invention is at least 2% greater (or at least 3% greater, or at least 4% greater, or at least 5% greater, or at least 6% greater, or at least 7% greater, or at least 8% greater, or at least 9% greater, or at least 10% greater) than a similar chromatography column assembly in which the average particle size of packing media within the guard column is equal to or less than the average particle size of packing media within the analytical column.
  • the present invention is also directed to methods of making chromatography column assemblies having unexpectedly superior system efficiency.
  • the method of making a chromatography column assembly comprises combining (i) an analytical column comprising a first packing media having a first average particle size with (ii) a guard column so that the guard column is in-line with the analytical column, wherein the guard column comprises a second packing media having a second average particle size that is greater than the first average particle size such that the resulting chromatography column assembly has a system efficiency greater than a similar chromatography column assembly in which the second average particle size is equal to or less than the first average particle size.
  • the above-described exemplary method of making a chromatography column assembly may include any number of additional steps. Suitable additional steps may include, but are not limited to, selecting at least one guard column and at least one analytical column having desired column dimensions; selecting packing media for the guard column and the analytical column based on possibly analyte(s) in a test sample; connecting the in-line guard column and analytical column to other assembly components such as those mentioned above (e.g., sample injector, detector, etc.); sealing one end of a given column; at least partially filling a column cavity of a given column with a rigid support material, such as any of the above- described rigid support materials; at least partially filling the column cavity of a given column with a first buffer solution to encapsulate the rigid support material; inserting a piston, spring and/or threaded rod into a given column to engage, and/or compress and/or retain the rigid support material within a desired area of the column; sealing an opposite end of a given column; and testing the pressure capacity of the resulting
  • the present invention is further directed to methods of using chromatography column assemblies.
  • chromatography column assemblies In one exemplary Docket No. W9795-01
  • the method of using a chromatography column assembly comprises a method of analyzing a test sample that potentially contains at least one analyte, wherein the method comprises the step of introducing the test sample into a chromatography column assembly comprising (i) an analytical column comprising a first packing media having a first average particle size, and (ii) a guard column in-line with the analytical column, the guard column comprising a second packing media having a second average particle size, the second average particle size being greater than the first average particle size; wherein the chromatography column assembly has a system efficiency greater than a similar chromatography column assembly in which the second average particle size is equal to or less than the first average particle size.
  • the method of analyzing a test sample may further comprise one or more of the following steps: changing packing media within the guard column and/or analytical column to improve resolution of analyte peak(s) on a chromatogram; adjusting the concentration of the test sample; changing the solvent(s) used in the mobile phase of the test sample; changing the column temperature; allowing the test sample to come into contact with the packing media; and allowing the test sample to remain in contact with the packing media for a desired period of time.
  • the test sample remains in contact with the packing media for a period of time ranging from about 1.0 minutes to about 10 minutes.
  • a VisionHT r T"M vl HPLC Column from Grace Davison Discovery Sciences (Columbia, MD), having dimensions 5.0 mm (length) x 2 mm (outer diameter), packed with particles having average particle sizes of 1.5 ⁇ m and 3.0 ⁇ m at a packing pressure of 1000 bars.
  • the particles were spherically-shaped silica-based inorganic particles having a particle surface area of about 200 m /g, and an average pore size of about 100 angstroms.
  • Table 2 below provides a comparison of sample guard columns comprising (1) particles having an average particle size of 1.5 ⁇ m, and (2) particles having an average particle size of 3.0 ⁇ m.

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  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Treatment Of Liquids With Adsorbents In General (AREA)
  • Solid-Sorbent Or Filter-Aiding Compositions (AREA)

Abstract

Cette invention se rapporte à des assemblages de colonnes de chromatographie. L'invention a également trait à des procédés de fabrication d'assemblages de colonnes de chromatographie et à des procédés d'utilisation d'assemblages de colonnes de chromatographie.
EP08795368A 2007-08-16 2008-08-15 Assemblages de colonnes de chromatographie Withdrawn EP2192963A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US96497007P 2007-08-16 2007-08-16
PCT/US2008/009784 WO2009023268A1 (fr) 2007-08-16 2008-08-15 Assemblages de colonnes de chromatographie

Publications (1)

Publication Number Publication Date
EP2192963A1 true EP2192963A1 (fr) 2010-06-09

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Application Number Title Priority Date Filing Date
EP08795368A Withdrawn EP2192963A1 (fr) 2007-08-16 2008-08-15 Assemblages de colonnes de chromatographie

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EP (1) EP2192963A1 (fr)
WO (1) WO2009023268A1 (fr)

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Publication number Priority date Publication date Assignee Title
CN103180727B (zh) * 2010-10-27 2015-12-16 通用电气健康护理生物科学股份公司 具有保护柱的色谱系统
EP3821238B1 (fr) * 2018-07-11 2022-08-31 Waters Technologies Corporation Systeme chromatogrphique et procede pour chromatographie en mode mixte piège-élution

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Publication number Priority date Publication date Assignee Title
GB693967A (en) * 1949-03-21 1953-07-08 California Research Corp Methods of separating mixtures of organic liquids by adsorption
CA1217654A (fr) * 1984-03-29 1987-02-10 Kurt E. Heikkila Analyse des additifs organiques dans les bains de galvanoplastie au moyen de nouvelles methodes chromatographiques
DE69817207T2 (de) * 1997-06-10 2004-06-17 Transgenomic, Inc., San Jose System und verfahren zur durchführung von polynukleotid-trennungen mit hilfe von flüssigchromatographie
WO1999034915A1 (fr) * 1998-01-06 1999-07-15 Cerus Corporation Dispositifs d'ecoulement permettant de reduire la concentration de composes dans des compositions biologiques et procedes associes
JP4879392B2 (ja) * 2000-11-09 2012-02-22 日本錬水株式会社 クロマト分離方法
US6679989B2 (en) * 2001-06-01 2004-01-20 Agilent Technologies, Inc. Integral, thru-bore, direct coupled high pressure liquid chromatography guard column

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WO2009023268A1 (fr) 2009-02-19

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