High throughput multi-dimensional sample analysis

• US 7,132,650 B1
• Filed: 09/25/2004
• Issued: 11/07/2006
• Est. Priority Date: 09/26/2003
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Nanostream, Inc.
• Inventors: Wilcox, Bruce, O'Connor, Stephen D., Gamble, Ronald C., Gregori, Matthew M.
• Primary Examiner(s): Wells, Nikita
• Assistant Examiner(s): Hashmi, Zia R.
• Application Number:
  US10/951,255
• Time in Patent Office: 773 Days
• Field of Search: None
• US Class Current: 250/288
• CPC Class Codes: G01N 2030/8447 Nebulising, aerosol formation or ionisation
• CPC Class Codes: G01N 30/466 with separation columns in parallel
• CPC Class Codes: G01N 30/7233 interfaced to liquid or supercritical fluid chromatograph interfaces in general for introducing or extracting samples to be analysed with specially adapted mass spectrometer, see H01J49/04
• CPC Class Codes: G01N 30/80 Fraction collectors
• CPC Class Codes: H01J 49/0018 Microminiaturised spectrometers, e.g. chip-integrated devices, Micro-Electro-Mechanical Systems [MEMS]
• CPC Class Codes: H01J 49/0431 for liquid samples interfaces to liquid chromatographs G01N30/7233
• CPC Class Codes: H01J 49/107 Arrangements for using several ion sources
• CPC Class Codes: Y10T 137/86863 Rotary valve unit
• CPC Class Codes: Y10T 137/86871 Plug
• CPC Class Codes: Y10T 436/115831 Condition or time responsive

Abstract

High throughput analytical systems and methods employing multiple liquid phase separation process regions coupled to a common mass spectrometer are provided. Disclosed systems and methods permits parallel separation and parallel storage of discrete eluate fractions, followed by sequential discharge and ionization of previously stored eluate portions to yield a composite ion stream containing the sequential series of eluate portions, followed by mass analysis of the ion stream. A common manifold may receive ions and utilize pressurized gas or ion gating to direct ions within the manifold toward the mass spectrometer inlet.

Claims

1. A high throughput analytical system comprising:

a plurality of liquid phase separation process regions adapted to operate in parallel;

a rotary storage device comprising a plurality of storage chambers, each storage chamber of the plurality of storage chambers being associated with and being in at least intermittent fluid communication with a different liquid phase separation process region of the plurality of liquid phase separation process regions;

a plurality of ionization sources disposed downstream of and in at least intermittent fluid communication with the plurality of storage chambers;

a mass spectrometer having a mass analyzer and an inlet in at least intermittent fluid communication with the plurality of ionization sources.

2. The system of claim 1 wherein the rotary storage device comprises a common actuator.

3. The system of claim 1 wherein the rotary storage device comprises a stationary portion and a rotating portion.

4. The system of claim 1 wherein each storage chamber of the plurality of storage chambers is microfluidic.

5. The system of claim 1, further comprising a manifold disposed between the plurality of ionization sources and the mass spectrometer inlet.

6. The system of claim 5, further comprising ion optics disposed within the manifold.

7. The system of claim 5, further comprising a supply of pressurized carrier gas in at least intermittent fluid communication with the manifold.

8. The system of claim 1, further comprising a supply of supplementing liquid in at least intermittent fluid communication with the plurality of ionization sources.

9. The system of claim 8 wherein each ionization source of the plurality of ionization sources operates in a substantially continuous manner.

10. The system of claim 1, further comprising a plurality of sensors disposed between the plurality of storage chambers and the plurality of ionization elements.

11. The system of claim 10 wherein the plurality of sensors comprises any of flow sensors and pressure sensors.

12. The system of claim 1, further comprising a supply of pressurized gas in intermittent fluid communication with each storage chamber of the plurality of storage chambers.

13. The system of claim 1 wherein the mass spectrometer comprises a time of flight mass spectrometer.

14. The system of claim 1 wherein the plurality of liquid phase separation process regions comprises at least ten liquid phase separation process regions.

15. The system of claim 1 wherein each liquid phase separation process region of the plurality of liquid phase separation process regions comprises a chromatography column.

16. The system of claim 1, further comprising a common source of mobile phase, wherein each liquid phase separation process region of the plurality of liquid phase separation process regions is in fluid communication with the common source of mobile phase.

17. The system of claim 1 wherein each ionization source of the plurality of ionization sources comprises an electrospray ionization source.

18. The system of claim 1 wherein each storage chamber has a fixed volume.

19. A high throughput analytical method comprising the steps of:

separating a plurality of samples in parallel utilizing a plurality of liquid phase separation process regions to yield a plurality of eluate streams;

storing a discrete fraction of each eluate stream of the plurality of eluate streams in parallel in a plurality of microfluidic storage regions; and

sequentially discharging and ionizing the discrete fraction of each eluate stream of the plurality of eluate streams to yield a composite ion stream comprising a series of ionized discrete fractions of each eluate stream of the plurality of eluate streams; and

analyzing the composite ion stream using a single mass spectrometer.

20. The method of claim 19, further comprising the steps of:

providing a plurality of ionization sources in at least intermittent fluid communication with the plurality of storage regions;

providing a manifold disposed between the plurality of ionization sources and the mass spectrometer; and

directing the ionized discrete fraction of each eluate stream within the manifold toward the mass spectrometer inlet.

21. The method of claim 20 wherein the directing step includes supplying a stream of pressurized gas to the manifold.

22. The method of claim 20 wherein the directing step includes generating at least one of an electric field and a magnetic field within the manifold.

23. The method of claim 19, further comprising the steps of:

providing a plurality of ionization sources in at least intermittent fluid communication with the plurality of storage regions; and

supplying a supplementing liquid to each ionization source of the plurality of ionization sources.

24. The method of claim 23 wherein the supplemental liquid supplying step is substantially uninterrupted during the sequential discharging step.

25. The method of claim 19, further comprising the step of sensing any of flow rate and pressure of the discrete fraction of each eluate stream of the plurality of eluate streams prior to the ionizing step.

26. The method of claim 25, wherein the ionizing step is responsive to the sensing step.

27. The method of claim 25, further comprising the steps of:

providing a plurality of ionization sources in at least intermittent fluid communication with the plurality of storage regions;

providing a manifold disposed between the plurality of ionization sources and the mass spectrometer; and

sequentially directing the ionized discrete fraction of each eluate stream within the manifold toward the mass spectrometer inlet, wherein the directing step is responsive to the sensing step.

28. The method of claim 19 wherein the sequential discharging step includes purging each storage region with a pressurized gas.

29. The method of claim 19, further comprising the step of detecting a property of each eluate stream of the plurality of eluate streams after the separating step, wherein the storing step is responsive to the detecting step.

30. The method of claim 19 wherein each liquid phase separation process region of the plurality of liquid phase separation process regions comprises a chromatography column, and the separating step includes performing a chromatographic separation process within each liquid phase separation process region of the plurality of liquid phase separation process regions.

31. The method of claim 19 wherein the plurality of microfluidic storage regions is disposed within a rotary device having a common actuator.

32. The method of claim 19 wherein the mass spectrometer comprises a time of flight mass spectrometer.

33. The method of claim 19 wherein each microfluidic storage region has a fixed first volume, and the discrete fraction of each eluate stream of the plurality of eluate streams has a volume substantially equal to the fixed first volume.

34. The method of claim 19 wherein the sequential discharge step includes discharging each microfluidic storage region of the plurality of microfluidic storage regions at least about once per second.

35. The method of claim 19 wherein the sequential discharge step includes discharging each microfluidic storage region of the plurality of microfluidic storage regions at least about twice per second.

36. The method of claim 19 wherein the sequential discharge step includes discharging each microfluidic storage region of the plurality of microfluidic storage regions at least about five times per second.

37. The method of claim 21 wherein the pressurized carrier gas is supplied to the manifold at a first flow rate, the previously stored eluate stream portions are supplied to the manifold at an aggregated second flow rate, and the first flow rate is at least about two times greater than the first flow rate.

38. The method of claim 21 wherein the pressurized carrier gas is supplied to the manifold at a first flow rate, the previously stored eluate stream portions are supplied to the manifold at an aggregated second flow rate, and the first flow rate is at least about five times greater than the first flow rate.

39. The method of claim 21 wherein the pressurized carrier gas is supplied to the manifold at a first flow rate, the previously stored eluate stream portions are supplied to the manifold at an aggregated second flow rate, and the first flow rate is at least about ten times greater than the first flow rate.

40. The method of claim 19 wherein:

the separating step includes generating a plurality of chromatograms for the plurality of samples;

each chromatogram of the plurality of chromatograms includes at least one sample component peak, with the plurality of chromatograms in the aggregate including a plurality of sample component peaks;

each sample component peak of the plurality of sample component peaks has a full-width/half-maximum peak duration;

the plurality of sample component peaks includes a peak having a minimum full-width/half-maximum peak duration; and

the sequential discharging and ionizing step is performed at least about three times within the minimum full-width/half maximum peak duration.

41. The method of claim 19 wherein:

the separating step includes generating a plurality of chromatograms for the plurality of samples;

each chromatogram of the plurality of chromatograms includes at least one sample component peak, with the plurality of chromatograms in the aggregate including a plurality of sample component peaks;

each sample component peak of the plurality of sample component peaks has a full-width/half-maximum peak duration;

the plurality of sample component peaks includes a peak having a minimum full-width/half-maximum peak duration; and

the sequential discharging and ionizing step is performed at least about five times within the minimum full-width/half maximum peak duration.

42. The method of claim 19 wherein:

the separating step includes generating a plurality of chromatograms for the plurality of samples;

each chromatogram of the plurality of chromatograms includes at least one sample component peak, with the plurality of chromatograms in the aggregate including a plurality of sample component peaks;

each sample component peak of the plurality of sample component peaks has a full-width/half-maximum peak duration;

the plurality of sample component peaks includes a peak having a minimum full-width/half-maximum peak duration; and

the sequential discharging and ionizing step is performed at least about ten times within the minimum full-width/half maximum peak duration.

43. A high throughput analytical method comprising the steps of:

providing a plurality of liquid phase separation process regions;

providing a plurality of microfluidic storage regions, each storage region being associated with and in at least intermittent fluid communication with a different liquid phase separation process region of the plurality of liquid phase separation process regions;

providing a plurality of ionization sources in intermittent fluid communication with the plurality of microfluidic storage regions;

providing a mass spectrometer having an inlet and having an associated manifold in at least intermittent fluid communication with the plurality of ionization sources;

supplying a plurality of samples to the plurality of liquid phase separation process regions;

separating the plurality of samples in parallel in the plurality of liquid phase separation process regions to yield a plurality of eluate streams;

storing a discrete fraction of each eluate stream of the plurality of eluate streams in parallel in a different microfluidic storage region of the plurality of microfluidic storage regions;

sequentially discharging and ionizing the discrete fraction of each eluate stream of the plurality of eluate streams into the manifold to yield a composite ion stream comprising a series of ionized discrete fractions of each eluate stream of the plurality of eluate streams; and

analyzing the composite ion stream using the mass spectrometer.

44. The method of claim 43, further comprising the step of supplying a stream of pressurized carrier gas to the manifold.

45. The method of claim 43, further comprising the step of sequentially directing the ionized discrete fraction of each eluate stream within the manifold toward the mass spectrometer inlet utilizing at least one of an electric field and a magnetic field.

46. The method of claim 43, further comprising the step of supplying a supplementing liquid to each ionization source of the plurality of ionization sources.

47. The method of claim 46 wherein the supplemental liquid supplying step is substantially uninterrupted during the sequential discharging step.

48. The method of claim 43, further comprising the step of sensing any of flow rate and pressure of the discrete fraction of each eluate stream of the plurality of eluate streams prior to the ionizing step.

49. The method of claim 48, wherein the ionizing step is responsive to the sensing step.

50. The method of claim 48, further comprising the step of sequentially directing the ionized discrete fraction of each eluate stream within the manifold toward the mass spectrometer inlet responsive to the sensing step.

51. The method of claim 43 wherein the sequential discharging step includes purging each storage region with a pressurized gas.

52. The method of claim 43, further comprising the step of detecting a property of each eluate stream of the plurality of eluate streams after the separating step, wherein the storing step is responsive to the detecting step.

53. The method of claim 43 wherein each liquid phase separation process region of the plurality of liquid phase separation process regions comprises a chromatography column, and the separating step includes performing a chromatographic separation process within each liquid phase separation process region of the plurality of liquid phase separation process regions.

54. The method of claim 43 wherein the plurality of microfluidic storage regions is disposed within a rotary device having a common actuator.

55. The method of claim 43 wherein the mass spectrometer comprises a time of flight mass spectrometer.

56. The method of claim 43 wherein each microfluidic storage region has a fixed first volume, and the discrete fraction of each eluate stream of the plurality of eluate streams has a volume substantially equal to the fixed first volume.

57. The method of claim 43 wherein the sequential discharge step includes discharging each microfluidic storage region of the plurality of microfluidic storage regions at least about once per second.

58. The method of claim 43 wherein the sequential discharge step includes discharging each microfluidic storage region of the plurality of microfluidic storage regions at least about twice per second.

59. The method of claim 43 wherein the sequential discharge step includes discharging each microfluidic storage region of the plurality of microfluidic storage regions at least about five times per second.

60. The method of claim 44 wherein the pressurized carrier gas is supplied to the manifold at a first flow rate, the previously stored eluate stream portions are supplied to the manifold at an aggregated second flow rate, and the first flow rate is at least about two times greater than the first flow rate.

61. The method of claim 44 wherein the pressurized carrier gas is supplied to the manifold at a first flow rate, the previously stored eluate stream portions are supplied to the manifold at an aggregated second flow rate, and the first flow rate is at least about five times greater than the first flow rate.

62. The method of claim 44 wherein the pressurized carrier gas is supplied to the manifold at a first flow rate, the previously stored eluate stream portions are supplied to the manifold at an aggregated second flow rate, and the first flow rate is at least about ten times greater than the first flow rate.

63. The method of claim 43 wherein:

the separating step includes generating a plurality of chromatograms for the plurality of samples;

each chromatogram of the plurality of chromatograms includes at least one sample component peak, with the plurality of chromatograms in the aggregate including a plurality of sample component peaks;

each sample component peak of the plurality of sample component peaks has a full-width/half-maximum peak duration;

the plurality of sample component peaks includes a peak having a minimum full-width/half-maximum peak duration; and

the sequential discharging and ionizing step is performed at least about three times within the minimum full-width/half maximum peak duration.

64. The method of claim 43 wherein:

the separating step includes generating a plurality of chromatograms for the plurality of samples;

each chromatogram of the plurality of chromatograms includes at least one sample component peak, with the plurality of chromatograms in the aggregate including a plurality of sample component peaks;

each sample component peak of the plurality of sample component peaks has a full-width/half-maximum peak duration;

the plurality of sample component peaks includes a peak having a minimum full-width/half-maximum peak duration; and

the sequential discharging and ionizing step is performed at least about five times within the minimum full-width/half maximum.

65. The method of claim 43 wherein:

the separating step includes generating a plurality of chromatograms for the plurality of samples;

each chromatogram of the plurality of chromatograms includes at least one sample component peak, with the plurality of chromatograms in the aggregate including a plurality of sample component peaks;

each sample component peak of the plurality of sample component peaks has a full-width/half-maximum peak duration;

the plurality of sample component peaks includes a peak having a minimum full-width/half-maximum peak duration; and

the sequential discharging and ionizing step is performed at least about ten times within the minimum full-width/half maximum peak duration.