Unified test means for ion determination

• US 4,645,744 A
• Filed: 07/02/1985
• Issued: 02/24/1987
• Est. Priority Date: 05/12/1983
• Status: Expired due to Term

• Current Assignee: Miles Laboratories Incorporated Elkhart Indiana A Corporation of
• Original Assignee: Miles Laboratories, Inc. (Bayer AG)
• Inventors: Hemmes, Paul, Lau, Arthur L. Y., Fleming, Roger L., Charlton, Steven C.
• Primary Examiner(s): NOT, DEFINED
• Application Number:
  US06/751,185
• Time in Patent Office: 602 Days
• Field of Search: 422/56, 422/57, 422/58, 436/73, 436/74, 436/79, 436/172, 436/175
• US Class Current: 436/74
• CPC Class Codes: C07C 37/50 by reactions decreasing the number of carbon atoms C07C37/04, C07C37/045, C07C37/055, C07C37/08 take precedence
• CPC Class Codes: C07C 39/14 with at least one hydroxy group on a condensed ring system containing two rings
• CPC Class Codes: C07C 45/46 Friedel-Crafts reactions
• CPC Class Codes: C07C 45/68 by increase in the number of carbon atoms
• CPC Class Codes: C07C 49/67 having two rings, e.g. tetralones
• CPC Class Codes: C07C 49/747 containing six-membered aromatic rings
• CPC Class Codes: C07C 51/347 by reactions not involving formation of carboxyl groups
• CPC Class Codes: C07C 57/30 containing six-membered aromatic rings
• CPC Class Codes: C07C 59/84 containing six membered aromatic rings
• CPC Class Codes: C07D 323/00 Heterocyclic compounds containing more than two oxygen atoms as the only ring hetero atoms
• CPC Class Codes: C07D 498/08 Bridged systems
• CPC Class Codes: G01N 31/22 using chemical indicators G01N31/02 takes precedence
• CPC Class Codes: G01N 31/221 for investigating pH value
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials

Abstract

A test means, test device and method for use for determining the presence of an ion in a test sample are disclosed. The test means comprises a substantially nonpolar, nonporous carrier matrix incorporated with an ionophore capable of forming a complex with the specific ion and reporter substance capable of interacting with the complex of ionophore and ion to produce a detectable response. The test means disclosed is particularly useful for serum potassium determinations. The test means forms the reagent layer in a multilayer format particularly suited to whole blood electrolyte determinations.

Claims

1. A test means for determining the presence of an ion in an aqueous test sample, the test means comprising:

a reagent layer composed of a substantially nonpolar, nonporous carrier matrix incorporated with

(a) an ionophore capable of forming a complex with a specific ion to be determined, and

(b) a reporter substance capable of interacting with the complex of the ionophore and the ion to produce a detectable response.

2. The test means of claim 1 in which the nonpolar, nonporous carrier matrix is a polymer film.

3. The test means of claim 2 in which the polymer film includes a plasticizer.

4. The test means of claim 3 in which the reporter substance is formed by at least two precursor substances incorporated in the nonpolar, nonporous carrier matrix, which formation is caused by the interaction of the precursor substances with the complex of the ionophore and the ion.

5. The test means of claim 3 in which the polymer film is additionally incorporated with light scattering centers.

6. The test means of claim 5 in which the light scattering centers are composed of particles selected from the group consisting of titanium dioxide, aluminum oxide, magnesium oxide, calcium carbonate, barium sulfate, zinc oxide, lead oxide, talc and microcrystalline cellulose.

7. A test means for determining the presence of an ion in an aqueous test sample, the test means comprising a reagent layer composed of a substantially nonpolar, nonporous carrier matrix incorporated with

(a) an ionophore capable of forming a complex with a specific ion to be determined;

and

(b) a neutral reporter compound having a dissociable proton, which proton is capable of dissociating from the reporter upon interaction with the complex of the ionophore and the ion to produce a detectable response.

8. The test means of claim 7, which additionally includes a water impervious support member affixed to the reagent layer.

9. The test means of claim 7 in which the nonpolar, nonporous carrier is a polymer film.

10. The test means of claim 9 in which the polymer film includes a plasticizer.

11. The test means of claim 7 in which the reporter substance is one capable of producing the appearance of, or change in, fluorescenee in the presence of the complex of the ionophore and the ion.

12. The test means of claim 11 in which the reporter substance is fluorescein or a derivative thereof.

13. The test means of claim 7 in which the reporter substance is one capable of producing the appearance of, or change in, color in the presence of the complex of the ionophore and the ion.

14. The test means of claim 13 in which the reporter substance is a compound having the structure:

##STR15## in which X, same or different, is a halogen or pseudohalogen;

in which each R, same or different, is a 2-, 3-, 5-, or 6-position substitutent, or multiple substituents thereof, selected from the lower alkyl, intermediate alkyl, aryl or fused ring at the 2,3- or 5,6-position; and

η

is 0 to 4.

15. The test means df claim 13 in which the reporter substance is a compound having the structure:

##STR16## in which R'"'"' is H or lower alkyl, R* is H or intermediate alkyl and X, same or different, is halogen or pseudohalogen.

16. The test means of claim 15 in which R'"'"' is methyl and R* is η

-decyl.

17. The test means of claim 16 in which the specific cation to be determined is potassium ion and the ionophore is composed of valinomycin and 2,3-naphtho-1,4,7,10,13-pentaoxacyclopentadeca-2-ene.

18. A multilayer test means for determining the presence of an ion in an aqueous test sample, comprising:

(a) a reagent layer comprising a substantially non-polar, nonporous carrier matrix incorporated with an ionophore capable of forming a complex with a specific ion to be determined and a reporter substance capable of interacting with the complex of the ionophore and the ion to produce a detectable response, and

(b) a reflecting layer disposed on top of the reagent layer and in laminar relationship thereto, said reflecting layer including one or more materials which act as a background in order to facilitate the detection of a detectable response in the reagent layer.

19. The process of determining unbound calcium, comprising the steps of:

(a) contacting the reflecting layer of the multilayer test means of claim 18 with an aqueous test sample; and

(b) determining the detectable response in the reagent layer.

20. The multilayer test means of claim 18 in which the specific ion to be determined is a cation and the reporter is a neutral compound having a dissociable proton, which proton is capable of dissociating from the reporter upon interaction of the reporter with the complex of the ionophore and the ion to produce a detectable response.

21. The multilayer test device of claim 18 which additionally includes a buffer incorporated with the reflecting layer.

22. The multilayer test device of claim 18 which additionally includes an interferant removal substance incorporated with the reflecting layer.

23. The multilayer test means of claim 18 in which the background material in the reflecting layer is composed of particles selected from the group consisting of titanium dioxide, aluminum oxide, magnesium oxide, barium sulfate, zinc oxide, lead oxide, talc and microcrystalline cellulose.

24. The multilayer test means of claim 23 in which the reflecting layer additionally includes a hydrophilic polymer.

25. The multilayer test means of claim 18 in which the nonpolar, nonporous carrier matrix is a polymer film.

26. The multilayer test means of claim 25 in which the polymer film includes a plasticizer.

27. The multilayer test means of claim 26 additionally comprising a transparent support member laminated to the reagent layer on the side opposed to the reflecting layer.

28. The multilayer test means of claim 18 additionally comprising an opacifying layer in laminar relationship to the reflecting layer and disposed on top of the reflecting layer, the opacifying layer containing particles for imparting an opaque appearance to the opacifying layer.

29. The multilayer test means of claim 28 in which the opacifying particles are particles of carbon black.

30. The multilayer test means of claim 28 in which the opacifying layer additionally includes a hydrophilic polymer.

31. The multilayer test device of claim 28 which additionally includes a buffer incorporated with the reflecting layer, the opacifying layer or both.

32. The multilayer test device of claim 28 which additionally includes an interferant removal substance incorporated with the reflecting layer, the opacifying layer or both.

33. A multilayer test means for determining the presence of an ion in a whole blood sample, comprising:

(a) a reagent layer comprising a substantially non-polar, nonporous carrier matrix incorporated with an ionophore capable of forming a complex with a specific ion to be determined and reporter substance capable of interacting with the complex of the ionophore and the ion to produce a detectable response, and

(b) a reflecting layer disposed on top of the reagent layer and in laminar relationship thereto, said reflecting layer including one or more materials wlich act as a background in order to facilitate the detection of a response in the reagent layer and sufficient sodium chloride to impart hematocrit independence to the ion determination.

34. A multilayer test means for determining the presence of an ion in a whole blood sample, comprising:

(a) a reagent layer comprising a substantially non-polar, nonporous carrier matrix incorporated with an ionophore capable of forming a complex with a specific ion to be determined and a reporter substance capable of interacting with the complex of the ionophore and the ion to produce a detactable response;

(b) a reflecting layer disposed on top of the reagent layer and in laminar relationship thereto, said reflecting layer including one or more materials which act as a background in order to facilitate the detection of a response in the reagent layer; and

(c) an opacifying layer in laminar relationship to the reflecting layer and disposed on top of the reflecting layer, the opacifying layer containing particles for imparting an opaque appearance to the opacifying layer;

wherein sufficient sodium chloride is incorporated in either the reflecting layer, the opacifying layer or both, to impart hematocrit independence to the ion determination.