TWO-DIMENSIONAL FLUID SEPARATION WITH CONTROLLED PRESSURE

• US 20150122655A1
• Filed: 05/10/2012
• Published: 05/07/2015
• Est. Priority Date: 05/10/2012
• Status: Active Grant

• Current Assignee: Agilent Technologies, Inc.
• Original Assignee: Agilent Technologies, Inc.
• Inventors: Choikhet, Konstantin, Witt, Klaus
• Granted Patent: US 9,694,301 B2
• Time in Patent Office: Days
• US Class Current: 204/601
• CPC Class Codes: B01D 15/18 relating to flow patterns
• CPC Class Codes: G01N 2030/322 pulse dampers
• CPC Class Codes: G01N 2030/328 valves, e.g. check valves of pumps
• CPC Class Codes: G01N 27/44704 Details; Accessories
• CPC Class Codes: G01N 30/32 of pressure or speed G01N30/36 takes precedence
• CPC Class Codes: G01N 30/34 of fluid composition, e.g. gradient G01N30/36 takes precedence
• CPC Class Codes: G01N 30/463 for multidimensional chromatography
• CPC Class Codes: G01N 30/465 with specially adapted interfaces between the columns
• CPC Class Codes: G01N 30/6095 Micromachined or nanomachined, e.g. micro- or nanosize
• CPC Class Codes: G01N 30/72 Mass spectrometers mass spectrometers per se H01J49/00

Abstract

A sample separation apparatus (200) for separating a fluidic sample, the sample separation apparatus (200) comprising a first separation unit (204) for separating the fluidic sample, a first fluid drive (202) configured for conducting the fluidic sample to be separated through the first separation unit (204), a second separation unit (208), arranged downstream of the first separation unit (204), for further separating the fluidic sample after treatment by the first separation unit (204), a second fluid drive (206) configured for at least partially conducting the fluidic sample, after treatment by the first separation unit (204), through the second separation unit (208), and a fluidic valve (218) having fluidic interfaces (222, 224, 226, 228) fluidically coupled to the first fluid drive (202) and the second fluid drive (206) and being switchable for performing the separation of the fluidic sample, wherein the sample separation apparatus (200) is configured for adjusting a pressure at a predefined position to a predefined value, wherein the predefined position is in a fluidic path between an outlet (270) of the first separation unit (204) and an inlet (272) of the second separation unit (208) or in fluid communication with this fluidic path.

Claims

1. A sample separation apparatus for separating a fluidic sample, the sample separation apparatus comprising

a first separation unit for separating the fluidic sample;

a first fluid drive configured for generating a fluid flow for conducting the fluidic sample to be separated through the first separation unit;

a second separation unit, arranged downstream of the first separation unit, for further separating the fluidic sample or fractions thereof after treatment by the first separation unit;

a second fluid drive configured for generating a fluid flow for conducting at least part of the fluidic sample after treatment by the first separation unit, through the second separation unit;

a fluidic valve having fluidic interfaces fluidically coupled to the first fluid drive, particularly fluidically coupled to the first fluid drive via the first separation unit, and fluidically coupled to the second fluid drive and being switchable for performing the separation of the fluidic sample;

wherein the sample separation apparatus is configured for adjusting, a pressure at a predefined position to a predefined value, wherein the predefined position is in a fluidic path between an outlet of the first separation unit and an inlet of the second separation unit or in fluid communication with this fluidic path.

2. The sample separation apparatus according to claim 1, wherein the sample separation apparatus is configured for adjusting the pressure at the predefined position to a constant value.

3. The sample separation apparatus according to claim 1, wherein the second fluid drive is controllable for adjusting the pressure at the predefined position to the predefined value.

4. The sample separation apparatus according to claim 1, wherein the second fluid drive is controllable for adjusting the pressure at the predefined position to follow a predefined course.

5. The sample separation apparatus according to claim 1, wherein the second fluid drive is controllable for adjusting a flow rate at the predefined position to a previously calibrated value.

6. The sample separation apparatus according to claim 1 wherein at least one of the first fluid drive and the second fluid drive is controllable for adjusting a ratio between a flow rate at the predefined position and a flow rate generated by the first fluid drive to a predefined value, particularly to a constant value, more particularly to follow a predefined course.

7. The sample separation apparatus according to claim 1, wherein a flow rate generated by the second fluid drive is controllable for adjusting, the pressure at the predefined position to the predefined value.

8. The sample separation apparatus according to claim 1, wherein the sample separation apparatus is configured for regulating the pressure at the predefined position to the predefined value based on sensor data measured in the fluidic path and being indicative of an actual pressure value at the position.

9. The sample separation apparatus according to claim 1, wherein the sample separation apparatus is configured for adjusting the pressure at the predefined position being located downstream of the fluidic valve to the predefined value.

10. The sample separation apparatus according to claim 1, wherein the sample separation apparatus is configured for adjusting the pressure at the predefined position being located at or fluidically connected to an outlet of or downstream of the second fluid drive to the predefined value.

11. The sample separation apparatus according to claim 1, wherein the sample separation apparatus is configured for adjusting the pressure to the predefined value in the entire fluidic path between the first separation unit, the second fluid drive and the second separation unit.

12. The sample separation apparatus according to claim 1, wherein the sample separation apparatus is configured for operation in accordance with a volume-based control scheme, particularly the sample separation apparatus is configured for executing sample separation by controlling run volumes of fluid flowing, over at least one of the separation units whereas evaluation of the sample separation is performed in terms of retention volumes required for releasing fractions of the fluidic sample from at least one of the separation units.

13. The sample separation apparatus according to claim 1, wherein the sample separation apparatus is configured for separating the fluidic sample by modifying a flow rate through at least one of the first fluid drive and the second fluid drive.

14. The sample separation apparatus according to claim 1, comprising, a flow coupler, particularly integrated in the fluidic valve, the flow coupler having two fluid inlet terminals and a fluid outlet terminal in fluid communication with one another, the fluid outlet terminal being fluidically connectable to the second separation unit.

15. The sample separation apparatus according to claim 1, wherein the first fluid drive the second fluid drive are in fluid communication with one another in at least one, particularly in each, switching state of the fluidic valve.

16. The sample separation apparatus according to claim 1, wherein the first fluid drive and the second fluid drive are both in fluid communication with a flow joiner directly or via a check valve, in at least one, particularly in each switching state of the fluidic valve.

17. The sample separation apparatus according to claim 1, wherein the first separation unit is arranged between the first fluid drive and the fluidic interface of the fluidic valve which the fluidic valve fluidically couples with the first fluid drive.

18. The sample separation apparatus according to claim 14, wherein the second separation unit is directly fluidically coupled to the fluid outlet terminal of the flow couplet.

19. The sample separation apparatus according to claim 14, wherein the flow coupler is configured as one of the group consisting of a fluidic T-piece, a fluidic Y-piece, and a fluidic X-piece.

20. The sample separation apparatus according to claim 14, wherein the flow coupler comprises at least one check valve preventing fluid from flowing in a reversed direction in at least one of the terminals.

21. The sample separation apparatus according to claim 1, wherein the fluidic valve comprises a first valve member and a second valve member being movable, particularly being rotatable, relative to one another to thereby adjust a respective switching state of the fluidic valve and thereby a corresponding one of different operation modes of the sample separation apparatus.

22. The sample separation apparatus according to claim 21, wherein the first valve member comprises ports forming the fluidic interfaces, and the second valve member comprises grooves for fluidically coupling different ports depending on a switching state of the fluidic valve.

23. The sample separation apparatus according to claim 1, wherein the fluidic valve is configured to be switchable to a first state in which the fluidic interface fluidically coupled to the first fluid drive is in fluid communication via the fluidic valve with the fluidic interface fluidically coupled to one of the fluid inlet terminals, and in which the fluidic interface fluidically coupled to the second fluid drive is in fluid communication via the fluidic valve with the fluidic interface fluidically coupled to the other one of the fluid inlet terminals.

24. The sample separation apparatus according to claim 23, wherein the fluidic valve is configured to be switchable to a second state in which the fluidic interface fluidically coupled to the first fluid drive is in fluid communication via the fluidic valve with the fluidic interface fluidically coupled to the other one of the fluid inlet terminals, and in which the fluidic interface fluidically coupled to the second fluid drive is in fluid communication via the fluidic valve with the fluidic interface fluidically coupled to the one of the fluid inlet terminals.

25. The sample separation apparatus according to claim 1, wherein at least one of the first fluid drive and the second fluid drive fluid gradient pump.

26. The sample separation apparatus according to claim 1, wherein the fluidic valve is switchable so that pressure conditions in the first separation unit and in the second separation unit remain constant before and after switching.

27. The sample separation apparatus according to claim 1, comprising a detector for detecting the separated fluidic sample and being arranged downstream of the second separation unit.

28. The sample separation apparatus according to claim 1, comprising a sample injector for injecting the fluidic sample into a mobile phase and being arranged between the first fluid drive and the first separation unit.

29. The sample separation apparatus according to claim 1, wherein the first fluid drive is operable with a first flow rate being smaller than a second flow rate according to which the second fluid drive is operable.

30. The sample separation apparatus according to claim 29, wherein the second flow rate is at least five times, particularly is at least ten times, more particularly is at least fifty times, of the first flow rate.

31. The sample separation apparatus according to claim 14, wherein the fluidic valve is switchable for performing the separation of the fluidic sample so that the first fluid drive and the second fluid drive are in fluid communication with one another via the flow coupler in each switching state of the fluidic valve.

32. The sample separation apparatus according to claim 1, comprising a control device configured for:

controlling the first separation unit to execute a first separation within a measurement volume interval for separating the fluidic sample into a plurality of fractions;

controlling the second separation unit to execute a sequence of second separations within the measurement volume interval for further separating at least a part of the plurality of separated fractions.

33. The sample separation apparatus according to claim 32, wherein at least one of the first separation and the sequence of second separations relates to a chromatographic gradient run.

34. The sample separation apparatus according to claim 32, wherein at least one of the first separation and the sequence of second separations relates to an isocratic chromatographic run.

35. The sample separation apparatus according to claim 1, wherein the first separation unit and the second separation unit are configured so as to execute the respective sample separation in accordance with different separation criteria, particularly in accordance with at least partially but not completely orthogonal separation criteria or in accordance with completely orthogonal separation criteria, more particularly having different selectivity patterns to the relevant sample components.

36. The sample separation apparatus according to claim 1, wherein the first separation unit and the second separation unit are configured so as to execute the respective sample separation on identical separation media but with different operating conditions, particularly at least one of the group consisting of different solvents, different steepness of elution gradients, different separation unit temperatures, and different pressures, so that the separation criteria are partially but not completely orthogonal.

37. The sample separation apparatus according to claim 1, comprising a detector unit arranged downstream of the first separation unit and upstream of the second separation unit.

38. The sample separation apparatus according to claim 1, comprising a sample treatment unit configured for additionally treating the fluidic sample after treatment by the first separation unit and before being delivered into the second separation unit, particularly by at least one of the group consisting of addition of chemical reagents, chemical modification, chemical derivatization, reaction detection, catalytic transformation, irradiation, and heating.

39. The sample separation apparatus according to claim 1, comprising at least one of the following features:

the sample separation apparatus comprises a control device configured for controlling operation of at least one of the group consisting, of the first fluid drive, the second fluid drive and the fluid;

at least one of the first separation unit and the second separation unit is configured for performing a separation in accordance with one of the group consisting of liquid chromatography, supercritical-fluid chromatography, capillary electrochromatography, electrophoresis and gas chromatography;

the sample separation apparatus is configured as a two-dimensional liquid chromatography sample separation apparatus, particularly being a comprehensive two-dimensional liquid chromatography apparatus;

the sample separation apparatus is configured to analyze at least one physical, chemical and/or biological parameter of at least one compound of the fluidic sample;

the sample separation apparatus comprises at least one of the group consisting of a chromatography device, a liquid chromatography device, an HPLC device, a gas chromatography device, a capillary electrochromatography device, an electrophoresis device, a capillary electrophoresis device, a gel electrophoresis device, and a mass spectroscopy device;

the sample separation apparatus is configured for generating a fluid flow for conducting the fluidic sample with a high pressure;

the sample separation apparatus is configured for generating a fluid flow for conducting the fluidic sample with a pressure of at least 100 bar, particularly of at least 500 bar, more particularly of at least 1000 bar;

the sample separation apparatus is configured to conduct a liquid fluid;

the sample separation apparatus is configured as a microfluidic device;

the sample separation apparatus is configured as a nanofluidic device;

at least one of the group consisting of the first separation unit and the second separation unit is configured for retaining at least a part of components of the fluidic sample and for allowing other components of the fluidic sample to pass;

at least one of the group consisting of the first separation unit and the second separation unit comprises a separation column;

at least one of the group consisting of the first separation unit and the second separation unit comprises a chromatographic column;

at least a part of at least one of the group consisting of the first separation unit and the second separation unit is filled with a separating material.

40. A method of separating a fluidic sample, the method comprising conducting the fluidic sample to be separated through a first separation unit by a fluid flow generated by a first fluid drive;

conducting, after treatment by the first separation unit, at least one fraction of the fluidic sample through a second separation unit downstream of the first separation unit assisted by fluid flow generated by a second fluid drive;

switching a fluidic valve having fluidic interfaces fluidically coupled to the first fluid drive and the second fluid drive for performing sequential separations of the at least one fraction of the fluidic sample after treatment by the first separation unit;

adjusting a pressure at a predefined position to a predefined value, wherein the predefined position is in a fluidic path between an outlet of the first separation unit and an inlet of the second separation unit or in fluid communication with this fluidic path.

41. The method according to claim 40, wherein the pressure is adjusted at the predefined position by controlling the flow rate of the second fluid drive.

42. The method according to claim 40, wherein the pressure is adjusted at the predefined position even when a gradient program of the second fluid drive will change the viscosity of the fluid flowing through the second separation unit.

43. A software program or product, preferably stored on a data carrier, for executing a method according to claim 40, when run on a data processing system such as a computer.