Capillary electrophoresis method and apparatus for electric field uniformity and minimal dispersion of sample fractions

• US 5,512,158 A
• Filed: 02/28/1995
• Issued: 04/30/1996
• Est. Priority Date: 02/28/1995
• Status: Expired due to Fees

• Current Assignee: Hewlett-Packard Company (HP Inc.)
• Original Assignee: Hewlett-Packard Company (HP Inc.)
• Inventors: Cole, Wesley D.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US08/395,268
• Time in Patent Office: 427 Days
• Field of Search: 204/299 R, 204/182.8, 204/180.1
• US Class Current: 204/453
• CPC Class Codes: G01N 27/44704 Details; Accessories

Abstract

A capillary electrophoresis method and apparatus for reducing dispersion of sample fractions are disclosed. The capillary tube in which the electrophoresis is performed has been flared, at least at its sample entrance end, to remove sharp corners which contribute to aberrations in the electric field distribution in a radial direction and result in differential migration of molecules depending on their proximity to the sharp corners. The flared tube, in contrast to a conventional tube, provides a more uniform electric field for electrophoresis and reduces undesired dispersion of the samples.

Claims

1. A method of forming a capillary tube for use in capillary electrophoresis comprising:

selecting an entrance end portion of said capillary tube, said entrance end portion having an inside diameter, interior and exterior surface, and an end surface that is generally perpendicular to said interior and exterior surfaces;

removing material from said interior surface of said entrance end portion such that said entrance end portion has a smooth contour with an increasing inside diameter with approach to said end surface; and

removing material at an exit end portion of said capillary tube such that a smooth contour is defined from said interior surface to a second end surface opposite to said end surface of said entrance end portion.

2. The method of claim 1 wherein removing said material is a step of removing material at the inside diameter of said capillary tube such that said interior surface is at an obtuse angle to said end surface.

3. The method of claim 1 wherein removing said material is a step of removing material at the inside diameter of said capillary tube such that the inside diameter at said end surface is increased by an amount in the range of two to three times.

4. The method of claim 1 wherein removing said material includes chemically etching said entrance end portion.

5. The method of claim 1 further comprising connecting said exit end portion to a second capillary tube having a similar contour.

6. A method for reducing dispersion of sample fractions during capillary electrophoresis, the method comprising:

providing a capillary tube with an entrance end, an exit end, and an internal diameter;

flaring the interior of the tube at the entrance end of the tube to remove sharp edges;

inserting a separation matrix into the tube;

inserting a sample for separation into the tube at the entrance end, the sample having electrically charged fractions;

placing each end of the tube in a buffer reservoir;

introducing an electrode into each buffer reservoir; and

applying an electric potential to the tube through the electrodes to create an electric field, the direction of the field being adjusted such that the sample will travel from the entrance end through the tube;

whereby the electric field is relatively uniform proximate to the entrance end.

7. The method of claim 6 wherein flaring the interior comprises etching the interior of the tube with a chemical.

8. The method of claim 7 wherein etching the interior with a chemical comprises treating the interior of the tube with hydrogen fluoride.

9. The method of claim 6 wherein flaring the interior comprises flaring so that the internal diameter of the tube is increased by an amount in the range of two to three times at the entrance end.

10. The method of claim 6 further comprising flaring the interior of the tube at the exit end of the tube to remove sharp edges.

11. The method of claim 10 further comprising aligning and coupling multiple tubes having flared entrance and exit ends in an end-to-end format such that the sample may be passed through the coupled tubes, whereby the electric field is relatively uniform throughout the coupled tubes.

12. An electrophoretic system for reducing dispersion of sample fractions, the system comprising:

a tube having an internal diameter, an entrance end and an exit end, the tube being flared at an interior at the entrance end, the interior having a smooth contour at a terminus of said entrance end;

at least two buffer reservoirs, each reservoir holding one of the entrance and exit ends of the tube; and

at least two electrodes, each electrode positioned substantially external to said tube in one of the reservoirs and connected to a source of electrical power to cause an electric field to pass a sample introduced at the entrance end of the tube toward the exit end of the tube.

13. The electrophoretic system of claim 12 wherein the tube is flared so that its internal diameter at said terminus is increased by an amount in the range of two to three times.

14. The electrophoretic system of claim 12 wherein the tube is flared at an interior at the exit end of the tube, the interior having a smooth contour at a terminus of said exit end.

15. The electrophoretic system of claim 14 further comprising multiple flared tubes aligned and coupled in an end-to-end format.

16. The electrophoretic system of claim 12 wherein the tube is flared by a chemical etching treatment with hydrogen fluoride.

17. An electrophoretic system for reducing dispersion of sample fractions, the system comprising:

a tube having an internal diameter, an entrance end and an exit end, the tube being flared at an interior at the entrance end and at the exit end, the interior having a smooth contour at a terminus of said entrance end;

at least two buffer reservoirs, each reservoir holding one of the entrance and exit ends of the tube; and

at least two electrodes, each electrode positioned in one of the reservoirs and connected to a source of electrical power to cause an electric field to pass a sample introduced at the entrance end of the tube toward the exit end of the tube.