Method and apparatus for NMR spectroscopy of nanoliter volume samples

• US 5,654,636 A
• Filed: 09/03/1996
• Issued: 08/05/1997
• Est. Priority Date: 11/14/1994
• Status: Expired due to Term

• Current Assignee: Board of Trustees of The University of Illinois
• Original Assignee: Board of Trustees of The University of Illinois
• Inventors: Wu, Nian, Sweedler, Jonathan V., Magin, Richard L., Peck, Timothy L., Webb, Andrew G.
• Primary Examiner(s): O'Shea, Sandra L
• Assistant Examiner(s): MAH, RAYMOND
• Application Number:
  US08/706,924
• Time in Patent Office: 336 Days
• Field of Search: 324/321, 324/300, 324/307, 324/318, 324/322, 422/70, 422/81, 436/161, 436/173
• US Class Current: 324/321
• CPC Class Codes: G01R 33/46 NMR spectroscopy

Abstract

A method and apparatus for structural elucidation and determination of a nanoliter size sample of analyte. The method includes separating the analyte using a microseparation technique and then substantially simultaneously analyzing or detecting the sample using a nuclear magnetic resonance (NMR) spectrometer. The apparatus includes an RF microcoil positioned for operable communication with a capillary member of the microseparation system and the NMR spectrometer to provide enhanced sensitivity and improved signal-to-noise ratio of the NMR spectrometer necessary to accurately analyze or detect nanoliter size samples.

Claims

1. A method of structural elucidation and determination of an analyte, comprising the steps of:

providing a sample of an analyte;

separating said analyte sample into nanoliter size (<

μ

L) samples; and

analyzing said nanoliter size samples with a nuclear magnetic resonance (NMR) spectrometer.

2. The method in accordance with claim 1 including providing a microcoil for operable communication with said NMR spectrometer to enhance the sensitivity of said NMR spectrometer.

3. The method in accordance with claim 1 wherein said sample size is less than 200 nanoliters.

4. The method in accordance with claim 1 wherein said step of analyzing said sample with said NMR spectrometer is accomplished in less than 5 minutes.

5. The method in accordance with claim 1 wherein said step of separating said sample is provided by capillary electrophoresis.

6. The method in accordance with claim 1 wherein said step of separating said sample is provided by liquid chromatography.

7. The method in accordance with claim 1 wherein said step of separating said sample is provided by capillary liquid chromatography.

8. An apparatus for structural elucidation and determination of an analyte, comprising:

microseparation means for separating said analyte into nanoliter size samples;

a nuclear magnetic resonance (NMR) spectrometer for measuring the nanoliter size samples provided by said means for separating; and

means for increasing the sensitivity of said NMR spectrometer to provide the sensitivity needed for effectively analyzing said nanoliter size samples.

9. The apparatus in accordance with claim 8 wherein said means for increasing the sensitivity of said NMR spectrometer includes an RF microcoil having a predetermined inside diameter selected for operable cooperation with said microseparation means and said NMR spectrometer, for providing enhanced sensitivity and improved signal-to-noise ratio of said NMR spectrometer, and for enabling said NMR spectrometer to analyze said nanoliter size samples of analyte.

10. The apparatus in accordance with claim 9 wherein said microseparation means are capillary electrophoresis.

11. The apparatus in accordance with claim 9 wherein said microseparation means are liquid chromatography.

12. The apparatus in accordance with claim 9 wherein said inside diameter of said RF microcoil is less than 1 mm.

13. The apparatus in accordance with claim 9 wherein said microseparation means include capillary electrophoresis means having a capillary member formed from fused silica, said microcoil being formed from wire wound about said capillary member in a detection region thereof.

14. The apparatus in accordance with claim 13 including a NMR probe in operable communication with said NMR spectrometer wherein said RF microcoil is positioned within said NMR probe substantially perpendicular to the magnetic field of said NMR spectrometer.

15. The apparatus in accordance with claim 9 wherein said microcoil is a multi-turn solenoidal coil.

16. The apparatus in accordance with claim 9 wherein said microcoil is a planar, lithographic coil.

17. The apparatus in accordance with claim 9 wherein said microcoil is fabricated onto a capillary or tubular structure forming said microseparation means through which said analyte flows.

18. The apparatus in accordance with claim 9 wherein said microcoil is fabricated onto a substrate forming said microseparation means across which said analyte flows.

19. The apparatus in accordance with claim 18 wherein said microcoil is fabricated onto a substrate onto which has been etched or grooved channels to serve as said capillaries through which said analytical compound flows.