Means for generating cell-disintegrated blood

Patent metadata

Field	Value
Publication number	US-9702864-B2
Application number	US-201414578136-A
Country	US
Kind code	B2
Filing date	Dec 19, 2014
Priority date	Dec 19, 2014
Publication date	Jul 11, 2017
Grant date	Jul 11, 2017

Abstract

A method for analyzing a whole blood sample can include injecting whole blood into a first zone of a dual zone sample loop, applying sufficient heat or energy to the whole blood to disintegrate the cellular components of the whole blood sample to produce cell disintegrated blood, and injecting a sufficient volume of the buffer into the dual zone sample loop to move the cell disintegrated blood into a second zone of the dual zone sample loop. The method can further include switching a multiport value to an inject position, flowing the cell disintegrated blood from the dual zone sample loop into a solid phase extraction column, and eluting components of the cell disintegrated blood from the solid phase extraction column into a liquid chromatography column.

First claim

What is claimed is:

1. A method of analyzing an intact whole blood sample, comprising:
sequentially loading a buffer, the intact whole blood sample, and an internal standard into a syringe assembly;
switching a multiport valve to a load position;
injecting the sequentially loaded internal standard and intact whole blood sample into a first zone of a dual zone sample loop;
applying sufficient energy to the intact whole blood sample within the first zone of the dual zone sample loop to disintegrate the cellular components of the intact whole blood sample to produce cell disintegrated blood;
injecting a sufficient volume of the buffer into the dual zone sample loop to move the cell disintegrated blood into a second zone of the dual zone sample loop;
switching the multiport value to an inject position;
flowing the cell disintegrated blood from the dual zone sample loop into a solid phase extraction column; and
eluting components of the cell disintegrated blood from the solid phase extraction column into a liquid chromatography column.

2. The method of claim 1 , further comprising eluting components of the cell disintegrated blood from the liquid chromatography column to a mass analyzer.

3. The method of claim 1 , further comprising identifying components of the cell disintegrated blood based on a retention time, a mass, a fragmentation mass, or any combination thereof.

4. The method of claim 1 , wherein the whole blood sample is resident in the first zone for a time sufficient to disintegrate the cellular components of the whole blood sample.

5. The method of claim 1 , wherein the whole blood sample moves through the first zone continuously, the flow rate and volume of the first zone resulting in a dwell time of the whole blood sample within the first zone sufficient to disintegrate the cellular components of the whole blood sample.

6. The method of claim 1 , wherein the whole blood sample stops within the first zone for a time sufficient to disintegrate the cellular components of the whole blood sample prior to injecting the buffer to move the cell disintegrated blood to the second zone.

7. A whole blood analysis system comprising:
an injection syringe assembly;
a multiport valve;
a dual zone sample loop comprising a first zone and a second zone;
a solid phase extraction column; and
a controller configured to:
sequentially load a buffer, an intact whole blood sample, and an internal standard into the syringe assembly;
switch the multiport valve to a load position;
inject the sequentially loaded internal standard and whole blood into a first zone of a dual zone sample loop;
applying sufficient heat to the intact whole blood sample within the first zone of the dual zone sample loop to disintegrate the cellular components of the whole blood sample to produce cell disintegrated blood;
inject a sufficient volume of the buffer into the dual zone sample loop to move the cell disintegrated blood into a second zone of the dual zone sample loop;
switch the multiport value to an inject position; and
flow the cell disintegrated blood from the dual zone sample loop into a solid phase extraction column to isolate a subset of components from the cell disintegrated blood.

8. The system of claim 7 , further comprising a liquid chromatography column.

9. The system of claim 8 , wherein the controller is further configured to flow elute the subset of components onto the liquid chromatography column.

10. The system of claim 9 , further comprising a mass analyzer.

11. The system of claim 10 , wherein the controller is further configured to elute components of the cell disintegrated blood from the liquid chromatography column to the mass analyzer.

12. The system of claim 11 , wherein the controller is further configured to identify components of the cell disintegrated blood based on a retention time, a mass, a fragmentation mass, or any combination thereof.

13. The system of claim 7 , wherein the controller is configured to move the whole blood sample through the first zone in a continuous flow, the flow rate and volume of the first zone resulting in a dwell time of the whole blood sample within the first zone sufficient to disintegrate the cellular components of the whole blood sample.

14. The system of claim 7 , wherein the controller is configured to stop the flow after injecting the whole blood sample into the first zone and prior to injecting the buffer, the flow being stopped while the whole blood sample is within the first zone and for a time sufficient to disintegrate the cellular components of the whole blood sample.

15. The system of claim 7 , wherein when the multiport valve is in the load position a fluid flow is from the syringe assembly into a first end of the dual zone sample loop adjacent to the first zone and out to waste from a second end of the dual zone sample loop adjacent to the second zone, and when the multiport value is in the inject position the fluid flow is from a pump into the first end of the dual zone sample loop and out to the solid phase extraction column from the second end of the dual zone sample loop.

Assignees

• Thermo Finnigan Llc

Inventors

• Brann John E
• Takarewski Jr Joseph J

Classifications

• G01N35/1016

  Control of the volume dispensed or introduced · CPC title

• G01N33/491

  by separating the blood components (G01N15/05 takes precedence) · CPC title

• G01N35/1097

  characterised by the valves (valves in general F16K) · CPC title

• G01N2035/00475

  Filters · CPC title

• G01N33/492 Primary

  Determining multiple analytes · CPC title