Means for Generating Cell-Disintegrated Blood

Patent metadata

Field	Value
Publication number	US-2016178608-A1
Application number	US-201414578136-A
Country	US
Kind code	A1
Filing date	Dec 19, 2014
Priority date	Dec 19, 2014
Publication date	Jun 23, 2016
Grant date	—

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

1 . A method of analyzing a whole blood sample, comprising:
loading a buffer, the whole blood sample, and an internal standard into a syringe assembly; switching a multiport valve to a load position; injecting the internal standard and the whole blood sample into a first zone of a dual zone sample loop; applying sufficient energy to the whole blood sample to disintegrate the cellular components of the 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 I., 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 loading pump; a dual zone sample loop comprising a first zone and a second zone, the first zone operable to supply energy to a whole blood sample; and a solid phase extraction column.

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

9 . The system of claim 7 , further comprising a mass analyzer.

10 . The system of claim 7 , further comprising a controller configured to:
load a buffer and the whole blood sample into the syringe assembly; switch the multiport valve to a load position; flow the whole blood sample from the syringe assembly through the first zone of a dual zone sample loop into the second zone of the dual zone sample loop, wherein sufficient energy is applied to the whole blood sample to disintegrate the cellular components of the whole blood sample to produce cell disintegrated blood; switch the multiport value to an inject position; and activate the loading pump to flow the cell disintegrated blood from the dual zone sample loop through a solid phase extraction column to separate a subset of components from the cell disintegrated blood.

11 . The system of claim 7 , wherein the multiport valve is configured to direct a fluid flow 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 when in a load position, and to direct a fluid flow from the loading pump into the first end and out to the solid phase extraction column from the second end when in an inject position.

12 . 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:
load a buffer, the whole blood sample, and an internal standard into the syringe assembly;
switch the multiport valve to a load position;
inject the internal standard and the whole blood into a first zone of a dual zone sample loop;
applying sufficient heat to the whole blood 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.

13 . The system of claim 12 , further comprising a liquid chromatography column.

14 . The system of claim 13 , wherein the controller is further configured to flow elute the subset of components onto the liquid chromatography column.

15 . The system of claim 14 , further comprising a mass analyzer.

16 . The system of claim 15 , wherein the controller is further configured to elute components of the cell disintegrated blood from the liquid chromatography column to the mass analyzer.

17 . The system of claim 16 , 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.

18 . The system of claim 12 , 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.

19 . The system claim 12 , 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.

20 . The system of claim 12 , wherein when the multiport value 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

• G01N2030/8822

  involving blood · CPC title

• G01N35/1016

  Control of the volume dispensed or introduced · CPC title

• G01N1/44

  Sample treatment involving radiation, e.g. heat · CPC title

• G01N30/88

  Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 · CPC title

• G01N30/7233

  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 title