ANALYSIS OF DRIED BLOOD SPOT SAMPLES IN A MICROFLUIDIC SYSTEM WITH DILUTION OF EXTRACTED SAMPLES

• US 20140208836A1
• Filed: 08/22/2012
• Published: 07/31/2014
• Est. Priority Date: 08/22/2011
• Status: Active Grant

• Current Assignee: Waters Technologies Corporation (Waters Corporation)
• Original Assignee: Waters Technologies Corporation (Waters Corporation)
• Inventors: Murphy, James P., Tomany, Michael J., Michienzi, Joseph D., Rainville, Paul, Plumb, Robert S., Gerhardt, Geoff C., Jung, Moon Chul
• Granted Patent: US 10,024,826 B2
• Time in Patent Office: Days
• US Class Current: 73/61.55
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0874 Three dimensional network
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: G01N 2001/383 collecting and diluting in a flow of liquid
• CPC Class Codes: G01N 2030/009 Extraction
• CPC Class Codes: G01N 2030/062 extracting sample from raw material
• CPC Class Codes: G01N 30/06 Preparation
• CPC Class Codes: G01N 30/16 Injection G01N30/24 takes precedence
• CPC Class Codes: G01N 30/6095 Micromachined or nanomachined, e.g. micro- or nanosize
• 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 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80

Abstract

An apparatus for use in a chromatography system includes a microfluidic substrate having a fluidic channel configured as an analytical chromatographic column and a fluidic port on one side of the microfluidic substrate. The fluidic port opens at a head end of the analytical chromatographic column. A dried blood spot (DBS) collection device holds one or more dried biological samples. The DBS collection device is directly coupled to the microfluidic substrate whereby one of the biological samples is placed into fluidic communication with the fluidic channel of the microfluidic substrate and an extraction of that biological sample flows toward the head end of the analytical chromatographic column. A diluent source fluidically coupled to the fluidic port supplies a solvent to the head end of the analytical column to dilute the extracted biological sample before the biological sample flows into the analytical chromatographic column.

Claims

1. A chromatography apparatus comprising:

a microfluidic substrate having a fluidic channel configured as an analytical chromatographic column and a fluidic port on one side of the microfluidic substrate, the fluidic port opening at a head end of the analytical chromatographic column;

a dried blood spot (DBS) collection device holding one or more dried biological samples, the DBS collection device being directly coupled to the microfluidic substrate whereby one of the biological samples is placed into fluidic communication with the fluidic channel of the microfluidic substrate and an extraction of that biological sample flows toward the head end of the analytical chromatographic column; and

a diluent source fluidically coupled to the fluidic port, the diluent source supplying a solvent to the head end of the analytical column to dilute the extracted biological sample before the biological sample flows into the analytical chromatographic column

2. The chromatography apparatus of claim 1, wherein the microfluidic substrate has a second fluidic channel configured as a sample-preparation column and a second fluidic port opening into the sample-preparation column, the sample-preparation column terminating at the head end of the analytical column, the DBS collection device coupling directly onto the microfluidic substrate such that the second fluidic port of the microfluidic substrate receives the extracted biological sample from the DBS collection device and passes the extracted biological sample into the sample-preparation column.

3. The chromatography apparatus of claim 2, wherein the sample-preparation column is configured for solid-phase extraction (SPE).

4. The chromatography apparatus of claim 1, further comprising a second microfluidic substrate having a second fluidic channel configured as a sample-preparation column, a first fluidic port on a first side of the second microfluidic substrate opening into one end of the sample-preparation column, and a second fluidic port on a second side of second microfluidic substrate,

wherein the DBS collection device couples directly onto the second microfluidic substrate such that the first fluidic port of the second microfluidic substrate receives the extracted biological sample from the DBS collection device and passes the extracted biological sample into the sample-preparation column, and

wherein the second side of the second microfluidic substrate is coupled to the other microfluidic substrate such that the second fluidic port of the second microfluidic substrate is in fluidic communication with a second fluidic port of the other microfluidic substrate and produces a fluidic path comprised of the sample-preparation and analytical chromatographic columns

5. The chromatography apparatus of claim 4, wherein the second microfluidic substrate has a plurality of fluidic channels configured as sample-preparation columns, the DBS collection device coupled directly onto the second microfluidic substrate has a different dried biological sample in fluidic communication with a different fluidic channel of the second microfluidic substrate, and the DBS collection device and the second microfluidic substrate are movably coupled to the other microfluidic substrate with the analytical chromatographic channel.

6. The chromatography apparatus of claim 5, further comprising an autosampler device mechanically coupled to the DBS collection device and to the second microfluidic substrate for indexing a different one of the sample-preparation columns of the second microfluidic substrate into alignment with the analytical column of the other microfluidic substrate.

7. The chromatography apparatus of claim 1, further comprising a pump fluidically coupled to one side of the DBS collection device to pump solvent to the dried biological sample in order to extract an analyte therefrom.

8. The chromatography apparatus of claim 7, wherein the pumped solvent is a supercritical fluid.

9. The chromatography apparatus of claim 7, wherein the pumped solvent is an organic solvent.

10. The chromatography apparatus of claim 7, wherein the pumped solvent is passes through the biological sample in order to extract the analyte.

11. The chromatography apparatus of claim 7, wherein the pumped solvent is flows atop the biological sample in order to extract the analyte.

12. The chromatography apparatus of claim 1, wherein the analytical chromatographic column has a 300 μ

m inner diameter.

13. The chromatography apparatus of claim 1, wherein the DB S collection device is a DBS card containing a plurality of dried biological samples.

14. The chromatography apparatus of claim 1, wherein the extraction and analysis of the biological sample occurs on-line.

15. The chromatography apparatus of claim 1, further comprising a microfluidic cartridge assembly housing the microfluidic substrate and the DBS collection device.

16. A method of separating a biological sample into constituent components, the method comprising:

coupling a dried blood spot (DBS) collection device, holding at least one dried biological sample, directly to a microfluidic substrate;

extracting an analyte from the dried biological sample and passing the extracted biological sample into a fluidic channel of the microfluidic substrate;

diluting the extracted biological sample in the fluidic channel before the extracted biological sample reaches an analytical chromatographic column; and

separating, by the analytical chromatographic column, the diluted biological sample into constituent components.

17. The method of claim 16, wherein the fluidic channel is configured as a sample-preparation column

18. The method of claim 17, wherein the sample-preparation column is configured for solid-phase extraction (SPE).

19. The method of claim 16, wherein the microfluidic substrate is a first microfluidic substrate, and further comprising coupling a second microfluidic substrate, having a fluidic channel configured as the analytical chromatography column, to the first microfluidic substrate to receive therefrom the diluted biological sample.

20. The method of claim 19, wherein the first microfluidic substrate has a plurality of fluidic channels configured as sample-preparation columns, the DBS collection device has a different dried biological sample is in fluidic communication with a different fluidic channel of the first microfluidic substrate, and the DBS collection device and the first microfluidic substrate are movably coupled to the other microfluidic substrate with the analytical chromatographic channel, and further comprising indexing a different one of the sample-preparation columns of the first microfluidic substrate into fluidic communication with the analytical chromatographic column of the second microfluidic substrate.

21. The method of claim 16, further comprising pumping solvent to the dried biological sample in order to extract an analyte therefrom.

22. The method of claim 21, wherein the pumped solvent is a supercritical fluid.

23. The method of claim 21, wherein the pumped solvent is an organic solvent.

24. The method of claim 23, further comprising passing the pumped solvent through the dried biological sample in order to extract the analyte.

25. The method of claim 23, further comprising passing the pumped solvent over atop the dried biological sample in order to extract the analyte.

26. The method of claim 16, wherein the analytical chromatographic column has a 300 μ

m inner diameter.

27. The method of claim 16, wherein the DBS collection device is a DBS card containing a plurality of dried biological samples.

28. The method of claim 16, wherein the extracting of the biological sample from the DB S collection device and separating the constituent components of the extracted biological sample occurs on-line.