Liquid chromatographic method and system

• US 20040188333A1
• Filed: 04/06/2004
• Published: 09/30/2004
• Est. Priority Date: 02/27/2001
• Status: Active Grant

• Current Assignee: Teledyne Instruments Incorporated (Teledyne Technologies Incorporated)
• Original Assignee: Isco, Inc. (Teledyne Technologies Incorporated)
• Inventors: Allington, Robert W., Davison, Dale A., Blakley, Scott L.
• Granted Patent: US 7,195,461 B2
• Time in Patent Office: Days
• US Class Current: 210/198.2
• CPC Class Codes: B01D 15/12 relating to the preparation of the feed
• CPC Class Codes: B01D 15/166 Fluid composition conditioning, e.g. gradient
• CPC Class Codes: B01D 15/247 Fraction collectors
• CPC Class Codes: G01N 1/38 Diluting, dispersing or mixing samples
• CPC Class Codes: G01N 13/00 Investigating surface or boundary effects, e.g. wetting power; Investigating diffusion effects; Analysing materials by determining surface, boundary, or diffusion effects scanning-probe techniques or apparatus G01Q
• CPC Class Codes: G01N 2001/383 collecting and diluting in a flow of liquid
• CPC Class Codes: G01N 2030/027 Liquid chromatography
• CPC Class Codes: G01N 2030/324 speed, flow rate
• CPC Class Codes: G01N 2030/326 pumps
• CPC Class Codes: G01N 30/22 in high pressure liquid systems
• CPC Class Codes: G01N 30/24 Automatic injection systems
• 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 30/466 with separation columns in parallel
• CPC Class Codes: G01N 30/74 Optical detectors measurement of intensity, velocity, spectral content, polarisation, or phase of infrared, visible or ultraviolet light G01J
• CPC Class Codes: G01N 30/84 Preparation of the fraction to be distributed
• CPC Class Codes: G01N 30/88 Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
• CPC Class Codes: G01N 35/1097 characterised by the valves valves in general F16K

Abstract

To economically perform preparatory chromatography, a plurality of pumps each having a corresponding one of a plurality of pistons and a corresponding one of a plurality of cylinders are driven by one motor to draw and pump solvent simultaneously into corresponding columns. To form a gradient, the pumps are connected to two-way valves that are connected alternately to a first solvent and a second solvent, whereby the time said valve is in a first position controls the amount of solvent drawn from the first reservoir into said pumps and the amount of time in said second position controls the amount of said second solvent drawn from the second reservoir into said pumps and the solvent is mixed in the pumping systems. The detectors are photodiodes mounted to light guides in the flow cells that generate signals related to light absorbance and communicate with a controller, whereby the controller receives signals indicating solute between the light guides and causes collection of solute. An over-pressure system compensates for pressure over a predetermined level.

Claims

1. A pumping system comprising:

at least one pump having a cylinder, a piston and a pump head with an outlet;

at least one inlet tube having first and second ends communicating with the cylinder at one end and adapted to communicate with at least two sources of fluid at the other end;

said at least one inlet tube having a diameter and length shaped for flow; and

drive means for driving said piston with sufficient speed to cause turbulent mixing in said cylinder, wherein fluid from said at least two sources of fluid are mixed before being pumped from said outlet;

an over-pressure system for compensating for pressures in the fluid above a preset pressure.

2. A pumping system according to claim 1 wherein the over-pressure system reduces the rate of flow of a channel having the pressure above the preset pressure.

3. A pumping system according to claim 1 wherein the over-pressure system includes a subsystem that tests for a flow rate at which the pressure is less than a predetermined pressure.

4. A pumping system according to claim 3 in which said at least two syringe pumps are driven by at least one pump motor and the over-pressure system includes a subsystem that shuts the at least one pump motor off when the subsystem that tests for a flow rate does not select a flow rate at which said pressure is below the preset pressure.

5. A pumping system according to claim 1 in which the over-pressure system includes a subsystem that completes a pump cycle at a lower pumping rate by slowing the pumps upon detecting a pressure above a preset pressure and then restores the flow rate to its original rate.

6. A pumping system in accordance with claim 5 further including an electronically controlled valve having an outlet and at least two inlets communicating at said outlet with said second end of said tube and at a first of said at least two inlets with one source of fluid and at a second of said at least two inlets with a second source of fluid whereby said at least one pump may pump a mixture of fluids.

7. A pumping system in accordance with claim 6 further including control means for switching said valve outlet from one of said at least two inlets to the second of said at least two inlets at least once during a refill stroke of said at least one pump.

8. A pumping system in accordance with claim 7 further including at least one syringe pump wherein said at least one syringe pump has a piston and a cylinder;

said pump having a refill flow rate at least 3 times faster than its delivery flow.

9. A pumping system in accordance with claim 8 in which at least one time-proportioning electronically controllable liquid gradient switching valve is arranged to produce consecutive pulses of liquid from at least one of said sources of fluid to a refill inlet at a fluid velocity high enough to induce turbulent mixing in a space between a head of the piston and that part of the cylinder not occluded by the piston.

10. A pumping system in accordance with claim 9 wherein said at least one pump includes a plurality of pumps, said pumping system further including means for synchronizing the at least one time-proportioning electronically controllable liquid gradient switching valve with refill movement of the said piston so that one charge of each desired fluid at a desired volume proportion is deposited in each pump and mixed to form at least one part of a step of a stepped gradient.

11. A pumping system in accordance with claim 10 further including:

first means for shutting off fluid flow between said pump and the at least one time-proportioning electronically controllable liquid gradient switching valve during delivery;

second means for synchronizing the at least one time-proportioning electronically-controllable liquid gradient switching valve with refill movement of said piston so that one charge of each desired fluid at a desired volume proportion is deposited in each pump and mixed to form at least one part of a step of a stepped gradient; and

means for repeating the said first and second means at consecutively different or same fluid proportions to produce an entire stepped gradient.

12. A pumping system in accordance with claim 11 wherein at least two equal charges of each of two fluids are alternately delivered to an inlet of said at least one syringe pump;

said two fluids being mixed in the at least one time-proportioning electronically controllable liquid gradient switching valve during a refill stroke of said piston, and then delivered as a single step of a step gradient, said refill stroke being sufficiently rapid to cause mixing in the cylinder of the pump.

13. A pumping system in accordance with claim 12 wherein more than one consecutive, entire, synchronous piston cycle corresponds to a single step of the gradient.

14. A pumping system in accordance with claim 13 wherein the stepped gradient is defined by the steps of the gradient taken consecutively.

15. A pumping system in accordance with claim 14 wherein said plurality of pumps comprises a multiple channel liquid chromatographic system having a parallel moving frame attached to a corresponding plurality of pistons for said plurality of pumps, wherein the motion of each of the pistons with respect to a corresponding cylinder is carried out by the parallel moving frame.

16. A pumping system in accordance with claim 15 further including:

a data system;

a sample concentration detector having an electrical output;

said data system including a recorder having an electrical recording output connection;

a fraction collector having a plurality of containers and a timing cycle for depositing liquid in the containers; and

the data system being electrically connected to the electrical output of said sample concentration detector and to the electrical recording output connection wherein a container charge timing cycle of the fraction collector is stopped during pump refill and restarted and run during liquid delivery of the at least one syringe pump.

17. A pumping system in accordance with claim 16 further including a first mixing means and second mixing means wherein the first mixing means resides in a fluid flow path between said at least one time-proportioning electronically controllable liquid gradient switching valve and said at least one syringe pump inlet and the second mixing means resides in the cylinder of said at least one syringe pump downstream of an inlet of the at least one time-proportioning electronically controllable liquid gradient switching valve.

18. A pumping system in accordance with claim 17 wherein the fluid flow path between the said at least one time-proportioning electronically controllable liquid gradient switching valve and said at least one syringe pump inlet is a flow passageway sized to produce mixing in the said fluid flow path, and of length or volume enough to make each step of the gradient sufficiently flat and reproducible for a desired set of chromatographic separation processes.

19. A pumping system in accordance with claim 18 wherein the flow passageway has a volume of at least one-tenth that of a single charge, said flow producing good axial mixing and poor transverse mixing and an outlet of said flow passageway injecting into the pump cylinder where the flow undergoes enhanced transverse mixing and axial mixing.

20. A pumping system in accordance with claim 17 wherein said at least one pump includes at least two syringe pumps and at least two equal charges of each of two fluids are alternately delivered to an inlet of at least one of said at least two syringe pumps;

said two fluids being mixed in the at least one time-proportioning electronically controllable liquid gradient switching valve during a refill stroke of said piston, and then delivered as a single step of a step gradient, said refill stroke being sufficiently rapid to cause mixing in a cylinder of the pump.

21. The pumping system of claim 20 wherein more than one consecutive, entire, synchronous piston cycle correspond to a single step of the gradient.

22. A pumping system in accordance with claim 21 wherein the stepped gradient is defined by the steps of the gradient taken consecutively.

23. A pumping system in accordance with claim 22 wherein each of said at least two syringe pumps includes a piston and a cylinder, said pumping system including a parallel moving frame attached to at least two pistons, wherein movement of each of the pistons with respect to a corresponding cylinder is carried out by the parallel moving frame.