Method and apparatus for determination of analyte concentration

• US 5,750,029 A
• Filed: 02/13/1995
• Issued: 05/12/1998
• Est. Priority Date: 10/16/1992
• Status: Expired due to Fees

• Current Assignee: Teledyne Isco, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Suprex Corp (Teledyne Technologies Incorporated)
• Inventors: Houck, Raymond K., Williams, Glen P., Koebler, Douglas J., Levy, Joseph M., Danielson, Victor G.
• Primary Examiner(s): Therkorn, Ernest G.
• Application Number:
  US08/388,274
• Time in Patent Office: 1,184 Days
• Field of Search: 210/634, 210/511, 210/137
• US Class Current: 210/634
• CPC Class Codes: B01D 11/0203 with a supercritical fluid
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: G01N 2001/4061 Solvent extraction
• CPC Class Codes: G01N 2030/062 extracting sample from raw material
• CPC Class Codes: G01N 33/24 Earth materials G01N33/42 takes precedence

Abstract

The present invention pertains to an apparatus for gathering analyte. The apparatus comprises a pumping mechanism for pumping solvent at supercritical pressure. The apparatus also comprises an extraction mechanism which receives a first sample and at least a second sample separate and distinct from the first sample and extract analyte from them essentially simultaneously. The extraction mechanism is connected to the pump mechanism to receive solvent at supercritical pressure for extracting the analyte from the samples. The apparatus additionally comprises a collection mechanism for collecting analyte from the solvent after the analyte has been extracted from the samples. The collection mechanism receives the supercritical solvent having the analyte extracted from the samples from the extraction mechanism. The present invention pertains to a method for extracting analyte. The method comprises the steps of pumping solvent at a supercritical pressure. Then there is the step of extracting analyte from N samples essentially simultaneously with the solvent at a supercritical pressure, where N is greater than or equal to two and is an integer. The N samples are separate and distinct from each other. Then there is the step of collecting the analyte of the N samples from the solvent.

Claims

1. An apparatus for extracting analyte comprising:

a pump mechanism for pumping solvent at supercritical pressure;

an extraction mechanism which receives N samples and extracts analyte from the N samples simultaneously, where N≧

2 and is an integer, said N samples separate and distinct from each other, said extraction mechanism connected to the pump mechanism to deliver solvent at supercritical pressure for extracting the analyte from the samples, said extraction mechanism having a first valve mechanism which controls which sample receives solvent at supercritical pressure from the pump, only one sample receiving solvent from the pump at any time; and

a collection mechanism for collecting analyte from the solvent after the analyte has been extracted from the samples, said collection mechanism receiving the supercritical solvent having the analyte extracted from the samples from the extraction mechanism, said collection mechanism having a valving mechanism which directs the flow of solvent from the extraction mechanism and which stops the flow of solvent from the extraction mechanism so the solvent is held in the extraction mechanism to extract analyte, said valving mechanism connected to the extraction mechanism.

2. An apparatus as described in claim 1 wherein the extraction mechanism receives a first sample and at least a second sample separate and distinct from the first sample and extracts analyte from them essentially simultaneously.

3. An apparatus as described in claim 2 wherein the collection mechanism includes a restrictor connected to the extraction mechanism which receives the solvent with analyte at a supercritical pressure and converts it to a pressure of 10 atmospheres or less, and a collector in communication with the restrictor which collects analyte in the solvent.

4. An apparatus as described in claim 3 wherein the extraction mechanism includes a first extraction chamber and at least a second extraction chamber, each chamber connected to receive solvent at a supercritical pressure from the pumping mechanism, and connected to the restrictor to pass solvent to it;

a first extraction vessel which holds the first sample, said first extraction vessel disposed in said first extraction chamber, and a second extraction vessel which holds the second sample, said second extraction vessel disposed in said second extraction chamber.

5. An apparatus as described in claim 4 wherein the pumping mechanism includes a pump and a first valve which directs solvent from the pump to the extraction mechanism when the first valve is in a first state and shuts off the solvent from the pump to the extraction mechanism when the first valve is in a second state.

6. An apparatus as described in claim 5 wherein the extraction mechanism includes a second valve connected to the first valve to receive solvent from the first valve when the first valve is in the first state, said second valve directs solvent to the first extraction chamber when the second valve is in a first state and directs solvent to the second extraction chamber when the second valve is in a second state.

7. An apparatus as described in claim 6 wherein the valving mechanism includes a third valve connected to the restrictor and connected to the extraction mechanism, said third valve directing solvent from the extraction mechanism to the restrictor when the third valve is in a first state, and stopping solvent from flowing from the extraction mechanism when the third valve is in a second state.

8. An apparatus as described in claim 7 wherein the second valve receives solvent from the first extraction chamber and directs it to the third valve when the second valve is in the first state and receives solvent from the second extraction chamber and directs it to the third valve when the second valve is in the second state.

9. An apparatus as described in claim 8 wherein the collector includes a vial that collects the analyte and allows the solvent to disperse into the atmosphere or to vent.

10. An apparatus as described in claim 9 wherein the restrictor is a variable restrictor.

11. An apparatus as described in claim 10 wherein the extraction mechanism includes a first plunger assembly in moveable contact with the first extraction chamber which connects with the first extraction chamber and seals the first extraction vessel when analyte is extracted from the first sample, said first plunger assembly in fluidic communication with the second valve so solvent in the first extraction chamber can flow to the second valve through the first plunger assembly;

and a second plunger assembly in moveable contact with the second extraction chamber which connects with the second extraction chamber and seals the second extraction vessel when analyte is extracted from the second sample, said second plunger assembly in fluidic communication with the second valve so solvent in the first extraction chamber can flow to the second valve through the second plunger assembly.

12. An apparatus as described in claim 11 wherein the first and second plunger assembly each are comprised of a plunger, a spring mechanism in which the plunger is seated and a forcing mechanism which moves the plunger, said respective plunger penetrating into and sealing the respective extraction vessel when the forcing mechanism forces the respective plunger into the respective vessel, and the spring mechanism retracting the respective plunger from the respective extraction vessel when the forcing mechanism is deactivated.

13. A method for extracting analyte comprising the steps of:

pumping solvent at a supercritical pressure;

extracting analyte from N samples essentially simultaneously with the solvent at a supercritical pressure, where N≧

2 and is an integer, said N samples separate and distinct from each other;

passing the solvent to a restrictor which reduces the solvent pressure to below supercritical pressure after the solvent has passed through a valve which controls the flow of solvent from the N samples; and

collecting the analyte of the N samples from the solvent.

14. A method for extracting analyte comprising the steps of:

pumping solvent at a supercritical pressure;

extracting analyte from N samples essentially simultaneously with the solvent at a supercritical pressure, where N≧

2 and is an integer, said N samples separate and distinct from each other;

passing the solvent to a restrictor which reduces the solvent pressure to below supercritical pressure after the solvent has passed through a valve which controls the flow of solvent from the N samples;

adjusting the restrictor automatically to avoid clogging in the restrictor; and

collecting the analyte of the N samples from the solvent.

15. A method as described in claim 14 wherein the extracting step includes the step of extracting analyte from a first sample and at least a second sample separate and distinct from the first sample.

16. A method as described in claim 15 wherein the extraction step includes the steps of providing the solvent at the supercritical pressure to the first sample;

providing the supercritical solvent at the supercritical pressure to the second sample;

gathering analyte of the first sample with the solvent at the supercritical pressure;

gathering analyte of the second sample with the solvent at the supercritical pressure;

flowing the solvent at the supercritical pressure with analyte of the first sample from the first sample;

reducing the pressure of the solvent with analyte of the first sample to below 10 atm;

flowing solvent at the supercritical pressure with analyte of the second sample from the second sample; and

reducing the pressure of the solvent with analyte of the second sample below 10 atm.

17. A system for use with supercritical fluid extraction comprising:

means for extracting analyte from a sample with supercritical fluid;

means for collecting analyte from the fluid;

said collecting means in fluidic communication with the extracting means;

means for providing desorbing solvent to the collecting means, said desorbent providing means in fluidic communication with the collecting means;

an automated variable restrictor for controlling flow of fluid with analyte from the extracting means to the collecting means and reducing the pressure of the supercritical fluid from the extracting means to a pressure below supercritical pressure for the collecting means and for preventing clogging of the flow of fluid between the extracting means and the collecting means, said restrictor having a first port in fluidic communication with the extracting means, a second port in fluidic communication with the collecting means, and a third port in fluidic communication with the desorbing providing means; and

a computer for automatically controlling the moving means, said computer in communication with the moving means.

18. A system as described in claim 17 wherein the restrictor includes a restrictor having a needle member with a flat end and means for selectively moving the needle member such that the fluid with analyte is converted from a first supercritical pressure in the first port to a second pressure in the second port which is less than the first pressure and below a supercritical pressure.

19. A system as described in claim 18 wherein the selectively moving means moves the needle member based on flow rate of fluid between the first port and the second port.

20. A system as described in claim 19 wherein the supercritical fluid includes modifier concentrations as great as 80% and the moving means moves the needle member to allow the fluid to pass between the first and second ports without clogging.

21. A system as described in claim 20 wherein the needle member has a diameter of less than 1/16 of an inch.

22. A system as described in claim 21 including a seat member disposed in the first, said seat member having a length less than 0.3 inches, said seat member having an internal diameter less than 0.016 inches, said seat member made of poly-ether-ether-ketone.

23. A system as described in claim 22 having a total assembled length of less than 2.5 inches and having a total assembled width of less than 1.5 inches.

24. A system as described in claim 23 wherein the restrictor has a dead volume no greater than 0.86 microliters.

25. A system as described in claim 17 wherein the selectively moving means moves the needle member based on pressure of the fluid.

26. A method as described in claim 25 wherein the needle member is made of a material having a coefficient of thermal expansion less than 6.2×

10^-6 in./in.

27. A system as described in claim 26 wherein the material is tungsten, silicon nitride, stainless steel, monel, 17-4PH, nitaonic, quartz, mica or plastic.

28. A system as described in claim 27 wherein the first supercritical pressure is up to 1000 atm.

29. A system as described in claim 25 wherein the needle member is made of a material having a low thermal conductivity, less than 116 BTU ft./hr. ft².

30. A system as described in claim 17 wherein the restrictor restricts supercritical solvent up having water up to 100% by content.

31. An apparatus for extracting analyte comprising:

a pump mechanism for pumping solvent at supercritical pressure;

an extraction mechanism which receives a sample and is connected to the pump mechanism to receive the solvent at supercritical pressure from the pump mechanism for extracting analyte from the sample;

a collection mechanism for collecting analyte from the solvent after the analyte has been extracted from the sample, said collection mechanism receiving the supercritical solvent having the analyte extracted from the sample from the extraction mechanism;

a valving mechanism connecting the pumping mechanism and the collection mechanism with the extraction mechanism, said valving mechanism controlling the flow of the solvent to the extraction mechanism and providing solvent through the extraction mechanism at least twice in a static state and twice in the dynamic state; and

an automated variable restrictor connected to the valving mechanism which reduces the solvent to below supercritical pressure and automatically adjusts to prevent being clogged, said restrictor connected also to the collection mechanism.

32. An apparatus as described in claim 31 wherein the valving mechanism includes a first valve connected to the pump, said first valve directs solvent from the pump to the extraction mechanism when the first valve is in a first state and shuts off the solvent from the pump to the extraction mechanism when the first valve is in a second state;

and a third valve connected to the extraction mechanism and the collection mechanism, said third valve directing solvent from the extraction mechanism to the collection mechanism when the third valve is in a first state, and stopping solvent from flowing from the extraction mechanism when the third valve is in a second state.

33. An apparatus as described in claim 32 wherein the collection mechanism includes a restrictor for reducing the supercritical pressure of the solvent to below supercritical pressure, said restrictor connected to the third valve.

34. An apparatus as described in claim 31 including means for providing modifier to the supercritical solvent, said modifier providing means connected to the valving mechanism.

35. An apparatus as described in claim 34 wherein the modifier providing means includes a plurality of modifier pump mechanisms for providing a plurality of different modifiers connected to the valving mechanism, each pump mechanism providing a corresponding modifier to the supercritical solvent through the valving mechanism.

36. An apparatus as described in claim 35 wherein the plurality of modifier pumps are disposed in parallel and connected to the valving mechanisms to provide tertiary and quaternary supercritical solvent and modifier mixtures to the extraction mechanism.

37. An apparatus as described in claim 36 wherein the valving mechanism includes a modifier tee connected to the modifier mechanisms through which modifier is introduced to the supercritical solvent.

38. An apparatus as described in claim 37 wherein the valving mechanism includes a check valve disposed between the pumping mechanism and the tee, said check valve preventing modifiers from back flowing to the pumping mechanisms from the modifier providing means.

39. An apparatus as described in claim 38 wherein each modifier pumping mechanism includes a modifier supply having corresponding modifiers.

40. An apparatus as described in claim 39 including a computer controlling the modifier pumping mechanisms, said computer connected to the modifier pumping mechanisms.

41. An apparatus as described in claim 34 wherein the modifier providing means includes a multi-headed modifier pump mechanism to provide multicomponent modifier to the supercritical solvent, said multi-headed modifier pump mechanism connected with the valving mechanism.

42. An apparatus as described in claim 41 wherein the multi-headed modifier pump mechanism is a dual-headed, triple-headed or four-headed modifier pump having dual, triple or quadruple modifier supplies, respectively, connected to the respective pumps.

43. A method for gathering analyte comprising the steps of:

flowing solvent at a supercritical pressure to a sample;

holding the solvent at the sample for a predetermined time so analyte in the sample is gathered by the solvent;

flowing the solvent away from the sample;

reducing the supercritical pressure of the solvent to below supercritical pressure;

collecting analyte from the solvent;

flowing solvent at a supercritical pressure to the sample;

holding the solvent at the sample for a predetermined time so analyte in the sample is gathered by the solvent;

flowing the solvent away from the sample;

reducing the supercritical pressure of the solvent to below supercritical pressure; and

collecting analyte from the solvent.

44. A method as described in claim 43 wherein the flowing supercritical solvent step includes the step of mixing modifier into the solvent.

45. A method as described in claim 44 wherein the mixing step includes the steps of pumping a desired modifier of a plurality of modifiers into the solvent.

46. A method as described in claim 44 wherein the mixing step includes the step of pumping multicomponent modifier from a combination of modifiers into the solvent.

47. An apparatus for extracting analyte comprising:

a pump mechanism for pumping solvent at supercritical pressure;

an extraction mechanism which receives a sample, said extraction mechanism connected to the pump mechanism to receive the solvent at supercritical pressure from the pump mechanism for extracting analyte from the sample;

a collection mechanism for collecting analyte from the solvent after the analyte has been extracted from the sample, said collection mechanism receiving the supercritical solvent having the analyte extracted from the sample from the extraction mechanism; and

a purge mechanism for removing moisture from the sample, said purge mechanism connected to the extraction mechanism.

48. An apparatus as described in claim 47 wherein the purge mechanism includes an inert gas supply for providing inert gas to the extraction mechanism to displace moisture in the sample.

49. An apparatus as described in claim 48 wherein the purge mechanism includes a first valve mechanism disposed between and connected to the pumping mechanism and the extraction mechanism, said inert gas supply connected to the first valve mechanism, said first valve mechanism allowing inert gas from the inert gas supply to flow to the extraction mechanism and solvent from the pumping mechanism from flowing to the extraction mechanism when the first valve mechanism is in a first state, said first valve mechanism stopping the flow of inert gas from the inert gas supply to the extraction mechanism when the first valve mechanism is in a second state.

50. An apparatus as described in claim 49 wherein the extraction mechanism includes a heating mechanism which heats the sample a desired amount while the inert gas flows through the extraction mechanism having the sample.

51. An apparatus as described in claim 50 wherein the first valve mechanism includes a first valve connected to the pumping mechanism and the extraction mechanism, and a purge valve mechanism having an open and closed state connected to the first valve, said first valve in the first state flowing solvent to the extraction mechanism, said first valve in a second state stopping solvent from flowing to the extraction mechanism, said first valve in a third state flowing inert gas from the inert gas supply to the extraction mechanism when the purge valve mechanism is in the open state.

52. An apparatus as described in claim 51 wherein the purge valve mechanism includes a purge valve having an open and a closed state;

a purge tee connected to the purge valve and the first valve and a vent valve having an open and closed state connected to the tee, said inert gas from the inert gas supply flowing to the first valve when the purge valve is in an open state and the vent valve is in a closed state.

53. An apparatus as described in claim 52 wherein the heating mechanism heats the sample to over 80°

C.

54. An apparatus as described in claim 53 wherein the inert gas supply provides inert gas at a pressure between 20 and 300 psi.

55. An apparatus for extracting analyte comprising:

a pump mechanism for pumping solvent at supercritical pressure;

an extraction mechanism which receives a sample, said extraction mechanism connected to the pump mechanism to receive the solvent at supercritical pressure from the pump mechanism for extracting analyte from the sample;

a collection mechanism for collecting analyte from the solvent after the analyte has been extracted from the sample, said collection mechanism receiving the supercritical solvent having the analyte extracted from the sample from the extraction mechanism; and

a precooler system for cooling solvent for supercritical extraction prior to the solvent being pumped by the pumping mechanism, said precooler system connected to the pumping mechanism.

56. An apparatus as described in claim 55 wherein the precooler system comprises a super cooler assembly for cooling the solvent, said super cooler assembly connected to the pumping mechanism;

and

a pump head cooler assembly connected to the pumping mechanism for cooling the pumping mechanism during operation of the pumping mechanism.

57. An apparatus as described in claim 56 wherein the super cooler assembly includes at least one thermo electric cooler having a cold side which is in thermal contact with the solvent and a hot side having a fan which expels heat from the thermo electric cooler.

58. An apparatus as described in claim 57 wherein the precooler system includes a tube coil in thermal contact with a tube coil through which the solvent flows through the thermo electric cooler.

59. An apparatus as described in claim 58 wherein the pumping mechanism has a pump head and the pump head cooler assembly includes a heat sink, and a thermally conductive bracket in contact with the pump head through which heat from the pump head escapes to the heat sink.

60. An apparatus as described in claim 59 wherein the heat sink includes a first fan and a second fan.

61. An extraction mechanism for extracting analyte from a sample comprising:

an extraction vessel which holds a sample; and

a plunger assembly which holds the vessel and penetrates into the vessel to introduce and remove solvent from the vessel, said plunger assembly having a plunger which moves up and down to penetrate into and separate from the vessel; and

a variable restrictor in fluid connection with the extraction vessel.

62. An extraction mechanism as described in claim 61 wherein the plunger assembly includes a spring mechanism for placing the plunger in a stable position, said spring mechanism connected to the plunger.

63. An extraction mechanism as described in claim 62 wherein the plunger assembly includes a forcing mechanism which moves the plunger into the vessel, and the spring mechanism retracts the plunger from the vessel when the forcing mechanism is deactivated.

64. An extraction mechanism as described in claim 63 wherein the plunger assembly includes a plate connected to the plunger and against which the spring mechanism presses against, said plate connected to the forcing mechanism which pushes against the plate and compresses the spring mechanism so the plunger penetrates the vessel when the forcing mechanism is activated.

65. An extraction mechanism as described in claim 64 wherein the forcing mechanism includes a hydraulic system.

66. An extraction mechanism as described in claim 65 wherein the hydraulic system includes a hydraulic cylinder connected to the plate and a hydraulic pump which pumps hydraulic fluid to the hydraulic cylinder to expand the hydraulic cylinder and move the plate.

67. An extraction mechanism as described in claim 62 wherein the spring mechanism holds the plunger in the vessel.

68. An extraction mechanism as described in claim 67 including a plate in contact with the plunger and the spring mechanism, said spring mechanism pushes down on said plate causing said plunger to penetrate into the vessel.

69. An extraction mechanism as described in claim 61 including a plunger top extending perpendicularly from the plunger which a user lifts to insert the vessel below the plunger.

70. An apparatus for extracting analyte comprising:

a pump mechanism for pumping solvent at supercritical pressure;

an extraction mechanism which receives N samples and extracts analyte from the N samples essentially simultaneously, where N≧

2 and is an integer, said N samples separate and distinct from each other, said extraction mechanism connected to the pump mechanism to deliver solvent at supercritical pressure for extracting the analyte from the samples, and

a valving mechanism which controls the flow of solvent out of the extraction mechanism; and

an automated variable restrictor connected to the valving mechanism which reduces the solvent to below supercritical pressure and automatically adjusts to prevent being clogged.

71. An apparatus as described in claim 70 wherein the extraction mechanism includes an extraction chamber, said chamber connected to receive solvent at a supercritical pressure from the pumping mechanism and connected to the restrictor to pass solvent to it, a first extraction vessel which holds the first sample and a second extraction vessel which holds the second sample, said first and second vessels disposed in the extraction chamber.

72. An apparatus as described in claim 71 including means for heating the extraction chamber to heat the first and second vessels.

73. An extraction apparatus for supercritical fluid extraction comprising:

an extraction vessel having at least one opening through which supercritical fluid flows; and

a one-time use disposable seal dispersed over the one opening comprising a top layer having a hole for fluid to flow through, a middle layer which is porous in contact with the top layer, and a bottom layer having a hole for fluid to flow through in contact with the middle layer.

74. An apparatus as described in claim 73 wherein the top and bottom layer are each 0.02-0.03 inches thick and the middle layer is about 0.1 inches thick.

75. An apparatus as described in claim 74 wherein the top and bottom layers are made of acetal and the middle layer is made of filter paper.

76. A method for supercritical fluid extraction comprising the steps of:

placing a disposable seal comprising a top layer having a hole for fluid to flow through;

a middle layer which is porous in contact with the top layer; and

a bottom layer having a hole for fluid to flow through in contact with the middle layer over an opening of an extraction vessel, said extraction vessel having a sample in the extraction vessel;

flowing supercritical fluid through the extraction vessel and the seal so it passes through the hole of the top layer, then the porous middle layer and then the hole of the bottom layer; and

removing the disposable seal from the vessel.

77. A seal as described in claim 76 including after the removing step, there are the steps of placing another sample into the vessel, placing another disposable seal over the opening of the vessel;

and

flowing supercritical fluid through the extraction vessel and the other seal.