Chromatography column-electrophoresis system

• US 4,708,782 A
• Filed: 09/15/1986
• Issued: 11/24/1987
• Est. Priority Date: 09/15/1986
• Status: Expired due to Term

• Current Assignee: Sepragen Corporation
• Original Assignee: Sepragen Corporation
• Inventors: Andresen, Brian D., Saxena, Vinit
• Primary Examiner(s): Niebling, John F.
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US06/906,846
• Time in Patent Office: 435 Days
• Field of Search: 204/299 R, 204/180.1, 250/288, 250/288 R, 250/288 A, 210/198.2, 210/198.3, 210/656, 210/658, 210/659, 422/70, 436/161, 436/173
• US Class Current: 210/198.2
• CPC Class Codes: G01N 2030/386 Radial chromatography, i.e. with mobile phase traversing radially the stationary phase
• CPC Class Codes: G01N 27/44756 Apparatus specially adapted therefor
• CPC Class Codes: G01N 30/84 Preparation of the fraction to be distributed
• CPC Class Codes: Y10T 436/24 Nuclear magnetic resonance, electron spin resonance or other spin effects or mass spectrometry

Abstract

A chromatography column-electrophoresis system for directing the fluid output from the column directly to a point of use, further resolution or separation, detection and/or identification while conditioning or preparing the output for analysis by another instrument such as a mass spectrometer. The system provides for substantially simultaneous analysis in a mass spectrometer, for example, material separated initially by the column, followed by electrophoretic separation and resolution. The chromatography column utilizes horizontal or radial flow of the sample fluid through the separating medium. The column may be of a type having a quickly replaceable or disposable separating medium, or the entire column may be of a readily replaceable or disposable type. The electrophoresis apparatus is of a capillary tube type, one embodiment of which utilizes a pair of capillary tubes thereby providing for continuous operation.

Claims

1. A system for separating fluid components and conditioning at least certain of the components for analysis or other use, comprising the combination of:

a horizontal flow liquid chromatography column;

a capillary type electrophoresis apparatus; and

means for interconnecting said column with said electrophoresis apparatus and controlling flow of fluid components from said column into said electrophoresis apparatus;

said column being removably mounted on said means such that associated fluid components separated in said column are controllably deposited directly into said electrophoresis apparatus, said column including disposable fluid separation material, and a fluid distribution and component collection system in which associated fluid to be separated is adapted to be directed around at least a portion of one of side of the fluid separation material and directed radially through the fluid separation material with separated fluid components being collected at an opposite side of the fluid separation material for discharge into said means;

said electrophoresis apparatus including at least one capillary tube operatively connected to said means, a reservoir for electrically conducting buffer material operationally connected to said means, and means for connecting a positive terminal of a high voltage source on buffer material contained in said reservoir;

said means including a multi-port valve assembly operationally connected to said component collection system of said column to said capillary tube and to said buffer material reservoir, for selectively directing associated fluid components and associated buffer material into said capillary tube.

2. The combination of claim 1, said means additionally includes an externally threaded member secured to said valve assembly for threaded, removable interconnection with an internally threaded member on said column, and means extending through said threaded members for interconnecting said component collection system of said column with said valve assembly.

3. The combination of claim 2, wherein said externally threaded member comprises a first section secured to said valve assembly and having an outer surface, and a second section having a smaller cross-section than said first section ano protruding outwardly from said surface of said first section and provided with external threads, a fluid passageway extending through said first and second sections of said externally threaded member, said externally threaded protruding section of said member being connected to said internally threaded member of said column, said interconnecting means extending into said fluid passageway of said externally threaded member.

4. The combination of claim 3, wherein said internally threaded member of said column comprises a lower support member having a first section with one side thereof in abutment with the fluid separation material of said column and provided with a substantially centrally located opening, a second section of smaller cross-section than said first section and protruding outwardly from said first section and provided with a substantially centrally located threaded opening and an outer surface having a sealing means therein, said threaded opening of said section cooperating with said threaded protruding section of said externally threaded member such that wnen said threaded sections are threadedly interconnected, said outer surfaces of said members are in contact with eacn other and said sealing means preventing fluid leakage between said members, said interconnecting means extending through said openings in said lower support member and being connected at one end in said opening of said first section of said lower support member.

5. The combination of claim 4, wherein said opening of said first section of said lower support member is provided with threads, wherein said interconnecting means comprises a tube, and including threaded means for securing one end of said tube in said threaded opening of said first section.

6. The combination of claim 1, wherein said multi-port valve assembly includes:

a housing having a chamber therein,

a member movably located in said chamber of said housing, said member being provided with a passage extending therethrough,

said housing being provided with a plurality of ports extending from an outer surface to said chamber, one of said ports being operationally connected to said component collection system of said column, another of said ports being operationally connected to said capillary tube, and another of said ports being operationally connected to said buffer material reservoir,

whereby movement of said member in said housing chamber aligns said passage therein with said component collection system or with said capillary tube and said buffer material reservoir.

7. The combination of claim 6, wherein said means additionally includes a connector member secured to said housing of said multi-port valve assembly, said connector member including first and second sections, with said second section being of smaller cross-section than said first section and protruding outwardly from a surface of said first section, said protruding second section being provided with threads, and a fluid passageway extending through said first and second sections and aligned with said one of said ports of said valve assembly housing.

8. The combination of claim 7, wherein said column includes a support member having an opening therein in alignment with said fluid passageway of said connector member, said opening including a threaded section for threaded interconnection with said protruding second section of said connector member, said support member also including an outer surface adapted to contact said outer surface of said first section of said connector member, and sealing means positioned between said outer surfaces.

9. The combination of claim 8, additionally including interconnecting means between said component collection system of said column and said connector member, said interconnecting means being secured at one end of said opening in said support member and extending into said fluid passageway of said connector member.

10. The combination of claim 9, wherein said opening in said support member additionally includes another threaded section, wherein said interconnecting means comprises a tube, and wherein one end of said tube is secured by a threaded member in said another threaded section of said opening in said support member.

11. The combination of claim 1, wherein said separation material of said column is in the form of a wafer.

12. The combination of claim 1, wherein said separation material is positioned intermediate a pair of spaced porous frits.

13. The combination of claim 1, wherein said column is of a disposable type.

14. The combination of claim 1, wherein said electrophoresis apparatus includes a pair of capillary tubes, a first of said capillary tubes being operationally connected to said multi-port valve assembly, a second of said capillary tubes being connected to another reservoir for electrically conducting buffer material, and a power supply having a positive terminal thereof being connected to said first mentioned reservoir and a negative terminal thereof being connected to said another buffer reservoir.

15. The combination of claim 14, wherein said pair of capillary tubes are at least partially located in spaced relation within a casing, each of said capillary tubes extending into and terminating at one end in passageways in a support block positioned in said casing, said support block also having a groove extending between said passageways therein.

16. The combination of claim 15, additionally including a molecular sieve assembly positioned in said casing and closely adjacent said groove in said support block, and means for retaining said molecular sieve assembly in said casing

17. The combination of claim 15, wherein an end of said casing in which said molecular sieve is located is positioned within a chamber of a mass spectrometer, and additionally including means for removably securing said casing in said mass spectrometer in a vacuum tight arrangement.

18. The combination of claim 1, wherein at least a portion of said capillary tube is positioned within a casing and connected at one end to said multi-port valve assembly, another end of said capillary tube terminating adjacent an end of said casing, and means for electrically connecting said another end of said capillary tube to an electrical ground, said end of said casing being adapted to be located at a point of use of said electrophoresis apparatus.

19. The combination of claim 18, wherein said end of said casing is secured in a mass spectrometer in a vacuum tight relationship.

20. In a system consisting of a liquid chromatography column, an electrophoresis apparatus, and a mass spectrometer, the improvement comprising:

means for removably connecting at least one chromatography column directly to an electrophoresis apparatus through a valve assembly connected to at least one reservoir for containing electrically conducting buffer material and connected to a capillary tube of said electrophoresis apparatus, and

means for connecting said electrophoresis apparatus to said mass spectrometer in a vacuum tight relationship,

said chromatography column being of a horizontal flow type,

whereby movement of said valve assembly to a first location allows fluid components separated in said column to be directed through said connecting means into said valve assembly, and movement of said valve assembly to a second location allows buffer material from said reservoir to pass through said valve assembly and direct the fluid components therein into said capillary tube in which fluid components undergo conditioning by electrophoresis as the components and buffer material pass through the capillary tube.

21. The improvement of claim 20, wherein said chromatography column is of a disposable type.

22. The improvement of claim 21, wherein said chromatography column is constructed so as to be readily disassembled and contains a disposable separation medium.