Apparatus and control kit for analyzing blood sample values including hematocrit

• US 4,686,479 A
• Filed: 07/22/1985
• Issued: 08/11/1987
• Est. Priority Date: 07/22/1985
• Status: Expired due to Term

• Current Assignee: Nova Biomedical Corporation 200 Prospect Street Waltham MA
• Original Assignee: Nova Biomedical Corporation 200 Prospect Street Waltham MA
• Inventors: Coppola, Pat, Rodomista, Guy, MacIndoe, Robert, Fowler, James E., Young, Chung C., Coleman, Robert L., Grimes, John F., Spaziani, Fred, Baumeister, Ferdnand, Sullivan, Sheila M., Catalano, Lou
• Primary Examiner(s): Eisenzopf, Reinhard J.
• Assistant Examiner(s): Harvey, Jack B.
• Application Number:
  US06/757,573
• Time in Patent Office: 750 Days
• Field of Search: 324/439, 324/71.1, 324/71.4, 324/450, 204/1 T, 204/403, 204/406, 204/409, 204/411, 204/412, 204/416, 204/400, 204/418-420, 436/8, 436/70, 422/61, 422/68, 422/99, 73/864.86, 339/91 R, 339/255 R
• US Class Current: 205/781.5
• CPC Class Codes: G01N 33/4915 using flow cells flow cytometry G01N15/14

Abstract

Hematocrit level of a blood sample is measured by flowing the sample along a liquid flow path and using electrodes in the flow path to obtain electrical signals representative of the sample'"'"'s electrical conductivity and of the concentration of an ion species in the path, either before or after the sample measurement. Electrical signals are obtained for standardizing solution conductivity and ion species concentration. A tentative sample hematocrit value is derived from the sample and standardizing conductivity signals. Then the tentative hematocrit value is corrected with reference to the sample and standardizing ion concentration signals and to the known standardizing solution ion concentration value. A control solution of the ion species at known concentration is used for evaluating the hematocrit detection apparatus; the solution also includes an ion activity enhancing agent. A removable septum assembly is used in a blood sample component measuring apparatus in which at least two standardizing solutions are introduced into the flow path. The septum assembly comprises: (a) chamber-defining members, which define a plurality of chambers, the members being permanently attached to each other; and (b) flexible septa positioned between the chambers, each septum having a slit, the slits being aligned to sealably receive therethrough an elongated probe connected to a flush fluid and each remaining chamber being connected to a standardizing fluid introducing lines, the septum assembly being removably attached as a unit to the apparatus which includes means for releasably attaching the septum assembly in position to receive the probe through the slits.

Claims

1. A method for determining the hematocrit value of a blood sample by:

(a) providing apparatus comprising a liquid flow path, means in the flow path for generating an electrical signal representative of the electrical conductivity of liquid in the Path, and means in the flow path for obtaining an electrical signal representative of the concentration of at least one ion species in liquid in the flow path;

(b) introducing standardizing solution in the flow Path having a known concentration of said ion species and having a conductivity indicative of a known equivalent hematocrit value, and obtaining a signal representative of said standardizing solution conductivity and obtaining a signal representative of said known ion species concentration;

(c) either before or after introducing the standardizing solution, introducing the sample in the flow path and obtaining an electric signal representative of the sample conductivity and an electric signal representative of the sample ion-species concentration; and

(e) deriving a tentative sample hematocrit value responsive to the sample conductivity signal, with reference to said standardizing conductivity signal and to said known standardizing equivalent hematocrit value; and

(f) correcting said tentative sample hematocrit value with reference to said sample and standardizing ion concentration signals and to said known ion concentration value.

2. The method of claim 1 wherein said method further comprises providing, from time to time, an external validation of said apparatus by introducing a control solution in said flow path, said control solution having a known ion species concentration, and a conductivity representative of a known equivalent hematocrit level.

3. The method of claim 2 wherein said control solution equivalent hematocrit level is within a physiologically normal range, and said control solution ion species concentration is within a physiologically normal range.

4. The method of claim 2 wherein said control solution comprises an ion activity enhancing agent.

5. The method of claim 2 wherein said agent is a polyol.

6. The method of claim 5 wherein said polyol is selected from glycerol and polyalkyl glycols.

7. The method of claim 1 or claim 2 wherein said conductivity obtaining step comprises:

(a) providing electrodes in said flow path coupled to a constant amplitude AC circuit via a transformer;

(b) applying an AC signal to said electrodes from said circuit via said transformer; and

(c) detecting impedance reflected in said AC circuit.

8. The method of claim 1 or claim 2 wherein said method comprises performing the following steps in any order:

(a) obtaining said electrical signals representative of standardizing conductivity and standardizing ion concentration;

(b) storing signals representative of said known standardizing equivalent hematocrit value and said known standardizing ion concentration value;

(c) obtaining said electrical signals representative of sample ion concentration and sample condutivity;

(d) comparing said sample and said standardizing ion concentration signals with reference to said stored known standardizing concentration value signal to derive a signal representative of sample ion concentration value;

(e) comparing said sample and said standardizing conductivity signals with reference to said stored standardizing hematocrit value signal to derive a signal representative of a tentative sample hematocrit value;

(f) correcting said tentative sample hematocrit value signal with reference to said sample ion concentration signal and said stored standardizing ion concentration value signal.

9. The method of either claim 1 or claim 2 wherein said ion species is Na⁺ or Cl^-.

10. Apparatus for determining hematocrit value in a blood sample comprising:

(1) means for providing a fluid flow path;

(2) means in said flow path for providing an electrical signal representative of the conductivity of liquid passing along said flow path;

(3) means in said flow path for providing a signal representative of the concentration of an ion species in liquid passing along said flow path;

(4) means for introducing said blood sample into said flow path to obtain a signal representative of sample conductivity and of sample ion species concentration;

(5) means for introducing into said flow path a standardizing solution having a known concentration of an ion species and having a conductivity representative of a known equivalent hematocrit value;

(6) means for deriving a signal representative of a tentative sample hematocrit value from the sample conductivity signal, with reference to the standardizing conductivity signal and to the standardizing equivalent hematocrit value; and

(7) means for correcting said tentative sample hematocrit value with reference to said standardizing and sample ion concentration signals and to said known standardizing ion concentration.

11. The apparatus of claim 10 wherein said apparatus comprises means for storing either said sample or said standardizing conductivity signal, and means for comparing said conductivity signals with reference to said known standardizing equivalent hematocrit value to generate a signal representative of said tentative sample hematocrit value.

12. The apparatus of claim 11 wherein said means for correcting said tentative sample hematocrit value signal comprises means for storing either said standardizing or said sample ion concentration signal and comparing said concentration signals with reference to said known standardizing ion concentration value.

13. The apparatus of claim 10 wherein said apparatus comprises at least two standardizing solutions, each of which has a conductivity indicative of a known equivalent hematocrit value and a known ion concentration.

14. The apparatus of claim 10 wherein said ion species is Na⁺ or Cl^-.

15. The apparatus of claim 10 wherein said means for providing an electrical signal representative of conductivity comprises electrodes in said flow path, a constant amplitude AC circuit coupled to said electrodes via a transformer, and means for detecting reflected impedance in said AC circuit.

16. The apparatus of claim 15 wherein said means for providing a signal respresentative of conductivity comprises:

(1) a first transformer for coupling said AC circuit to said electrodes;

(2) a second transformer for maintaining constant amplitude in said AC circuit; and

(3) means establishing a loop, connected between said electrodes, comprising means connected in said loop between said electrodes and said second transformer to compensate for inherent capacitance at the electrode/sample interface.

17. The apparatus of claim 16 wherein said apparatus comprises an ion-species sensitive electrode positioned in said flow path and connected via an electrical circuit to the input of a multiplexer, said impedance detecting means also being connected to the input of said multiplexer, said multiplexer having an output means connected via an analog-to-digital converter to a means for storing and comparing signals, and to said means for correcting sample conductivity.

18. A control solution kit for evaluating apparatus that determines a tentative level for the hematocrit of a blood sample by determining the sample conductivity and correcting said tentative level with reference to a sample ion species concentration level, said kit comprising an aqueous solution comprising said ion species and an ion activity enhancing agent, said solution having a known concentration of said ion and a known equivalent hematocrit value.

19. The kit of claim 18 wherein said ion species is Na⁺ or Cl^-.

20. The kit of claim 19 wherein said agent is a polyol.

21. The kit of claim 19 wherein said polyol is selected from glycerol and polyalkyl glycols.

22. The kit of claim 18 wherein said ion concentration and said equivalent hematocrit level are within physiologically normal ranges.

23. Apparatus for determining the concentration of a blood sample component comprising means defining a flow path, means in said path for generating an electrical signal representative of said component concentration in fluids in said path, means for introducing at least two standardizing solutions in said flow path, and means for comparing the sample concentration signal with the standardizing concentration signals wherein said apparatus further comprises a septum assembly connected to said flow path means and to said standardizing fluid introducing means, said septum assembly comprising:

(a) chamber-defining members, said members defining a plurality of chambers, said members being permanently attached to each other; and

(b) flexible septa positioned between said chambers, each said septum having a slit, said slits being aligned to sealably receive therethrough an elongated probe connected to said flow path, each said chamber being connected to a said standardizing fluid introducing means, said septum assembly being removably attached as a unit to said apparatus, said apparatus including means for releasably attaching said septum assembly in position to receive said probe through said slits.

24. The apparatus of claim 23 wherein said releasable attaching means comprises a post on the base of said septum assembly and a means on said apparatus for cooperatively engaging said post, said engaging means comprising a pair of elongated resilient spring means spaced to receive and releasably engage said post therebetween.

25. The apparatus of claim 23 wherein each said chamber-defining members comprises an inlet means, and said standardizing fluid introducing means comprises an integral standardizing-fluid-flow-path manifold connector having a plurality of outlets, each of which is positioned and sized to removably seal to a said inlet means.

26. The apparatus of claim 25 wherein each said inlet means comprises a cylindrical inlet positioned in a recess in said chamber-defining member, and said manifold connector is cooperatively sized and shaped to fit within said recesses.

27. A septum assembly comprising means for removable attachment to apparatus for measuring the concentration of blood sample components, said assembly comprising chamber-defining members defining a plurality of chambers, said members being permanently attached to each other, and flexible septa positioned between said chambers, each said septa having a sealable slit, said slits being aligned, each said septum assembly chamber comprising means for connection to a standardizing fluid source, said septum assembly further comprising a means for releasable attachment as a unit to said analyzer in position to sealably receive through said aligned slits a hollow elongated analyzer probe.

28. The septum assembly of claim 27 wherein said releasable attachment means comprises a post on the base of said septum assembly configured to cooperatively engage a pair of elongated resilient spring means on said concentration measuring apparatus, spaced to receive and releasably engage said post therebetween.

29. The septum assembly of claim 27 wherein each said chamber-defining member comprises an inlet means and means surrounding said inlet means for receiving a manifold connector.

30. The septum assembly of claim 27 wherein each said septa comprises an annular ridge positioned radially outwardly of, and concentrically with, said slit to seal against a said chamber-defining member.