Analytical element and method for determination of total lacate dehydrogenase

• US 4,803,159 A
• Filed: 04/07/1986
• Issued: 02/07/1989
• Est. Priority Date: 06/03/1985
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Smith-Lewis, Margaret J.
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): Spiegel, Carol A.
• Application Number:
  US06/848,613
• Time in Patent Office: 1,037 Days
• Field of Search: 435/26, 435/805, 422/55, 422/56, 422/57, 422/58, 422/59, 436/169, 436/170, 436/175, 436/810
• US Class Current: 435/26
• CPC Class Codes: C12Q 1/32 involving dehydrogenase
• CPC Class Codes: G01N 33/526 the element being adapted for a specific analyte
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials

Abstract

A multilayer analytical element exhibits improved precision in the assay of total lactate dehydrogenase (LDH). The element also provides reduced susceptibility to total protein and hemoglobin interferences in the assay. The element contains a porous spreading layer composed of a particulate structure. A fluorinated surfactant is also used in the assay. This surfactant has one or more fluorocarbon moieties provided, that when the surfactant has more than one such moiety, they are substantially linear. The surfactant can be incorporated in the element or added at the time of the assay.

Claims

1. A dry analytical element for the determination of total lactate dehydrogenase (LDH) comprising a support having thereon

a porous spreading layer composed of a particulate structure comprising a plurality of particles being bonded to each other on surface areas of said particles which are adjacent, where said adjacent particles form a coherent, three dimensional lattice which is essentially non-swellable to an aqueous liquid,

said element further comprising a saturated or unsaturated fluorinated surfactant containing one or more fluorocarbon moieties each having from 4 to 16 carbon atoms and from 8 to 33 fluorine atoms, and each of said moieties having a ratio of fluorine atoms to carbon atoms of about 2:

1, provided that when said surfactant contains more than one such moiety, said moieties have up to two fluorinated or unfluorinated methyl branches.

2. The element of claim 1 further comprising a substrate for LDH wherein the molar ratio of said fluorinated surfactant to said substrate is from about 0.01:

1 to about 10;

1.

3. The element of claim 2 wherein the molor ratio of said fluorinated surfactant to said substrate is from about 0.25:

1 to about 2.5;

1.

4. The element of claim 1 further comprising a second layer containing an indicator material which provides a detectable spectrophotometric change in response to the reaction of LDH with said substrate.

5. The element of claim 1 wherein said fluorinated surfactant is located in said spreading layer.

6. The element of claim 1 wherein said surfactant has only one fluorocarbon moiety.

7. A dry multilayer analytical element for the determination of total lactate dehydrogenase (LDH), said element comprising a support having thereon, in order from said support and in fluid contact,

a registration layer comprising an indicator material which provides a detectable spectrophotometric change in response to the reaction of LDH with a substrate for LDH, and

an isotropically porous spreading layer comprising said LDH substrate and composed of a particulate structure comprising:

(i) a plurality of heat-stable, organo-polymeric particles non-swellable in and impermeable to an aqueous liquid, and having a particle size of from about 1 to about 200 μ

m, and

(ii) an adhesive, in an amount of up to about 10 weight percent of said particles, comprising an organic polymer different from that of said particles,

substantially all of said adhesive being concentrated on surface areas of adjacent particles where said adjacent particles are in closest proximity, and bonding said particles into a coherent, three-dimensional lattice which is essentially non-swellable in said liquid,

said element containing, in any of said layers, a saturated or unsaturated fluorinated surfactant containing one or more fluorocarbon moieties each having from 4 to 16 carbon atoms and from 8 to 33 fluorine atoms, and each of said moieties having a ratio of fluorine atoms to carbon atoms of about 2;

1, provided that when said surfactant contains more than one such moiety, said moieties have up to two fluorinated or unfluorinated methyl branches, which surfactant is present in a molar ratio to said substrate of from about 0.01;

1 to about 10;

1.

8. The element of claim 7 wherein said fluorinated surfactant is selected from the group consisting of a perfluoroalkyl quaternary ammonium salt and a perfluoroalkyl betaine.

9. The element of claim 7 wherein said fluorinated surfactant is located in said spreading layer.

10. The element of claim 7 wherein said particles comprise an addition polymer formed from one or more of the following ethylenically unsaturated polymerizable monomers:

(a) up to 100 weight percent of an amino-substituent-free vinyl carbocyclic aromatic,

(b) up to about 25 weight percent of an acrylic acid ester,

(c) up to 100 percent of a methacrylic acid ester,

(d) up to about 30 weight percent of an ethylenically unsaturated carboxylic acid

(e) up to about 75 weight percent of an ethylenically unsaturated nitrile,

(f) up to about 20 weight percent of an amino-substituted vinyl carbocyclic aromatic,

(g) up to about 20 weight percent of an ethylenically unsaturated crosslinkable monomer,

(h) up to about 20 weight percent of a tertiary aminoalkyl acrylate or methacrylate,

(i) up to 100 weight percent of an N-heterocyclic vinyl monomer, and

(j) up to about 20 weight percent of an acrylamide or methacrylamide,

and said adhesive comprises an addition polymer formed from ethylenically unsaturated polymerizable monomers selected from the following group;

A. A blend containing from about 1 to about 35 weight percent of one or more amino-substituent-free vinyl carobcyclic aromatics and from about 65 to about 99 weight percent of one or more alkyl acrylates or methacrylates,

B. a blend containing from about 20 to about 95 weight percent of one or more amino-substituent-free vinyl carbocyclic aromatics, acrylic or methacrylic acid esters and ethylenically unsaturated polymerizable crosslinkable monomers, and from about 5 to about 80 weight percent of one or more ethylenically unsaturated polymerizable monomers having an active hydrogen or salts thereof,

C. a blend containing from about 15 to 100 weight percent of one or more ethylenically unsaturated monomers selected from the group consisting of 1-vinylimidazole, N-vinyl-2-pyrrolidone, vinylbenzyl alcohol, ethyl acrylate or an acrylamide or methacrylamide, and up to 85 weight percent of one or more ethylenically unsaturated polymerizable crosslinkable monomers, and

D. a blend containing from about 80 to about 98 weight percent of one or more acrylic or methacrylic acid esters, and from about 2 to about 20 weight percent of one or more ethylenically unsaturated polymerizable monomers containing one or more anionic moieties.

11. The element of claim 7 wherein said substrate is pyruvic acid or a salt thereof, and said indicator material is nicotinamide adenine dinucleotide, reduced form.

12. A method for the determination of total lactate dehydrogenase (LDH) comprising the steps of:

A. in the presence of a substrate for LDH, an indicator material which provides a detectable change in response to the reaction of LDH with said substrate, and a saturated or unsaturated fluorinated surfactant containing one or more fluorocarbon moieties each having from 4 to 16 carbon atoms and from 8 to 33 fluorine atoms, and each of said moieties having a ratio of fluorine atoms to carbon atoms of about 2;

1,

provided that when said fluorinated surfactant contains more than one fluorocarbon moiety, said moieties have up to two fluorinated or unfluorinated methyl branches,

contacting an analytical element with a sample of a liquid suspected of containing LDH, said element comprising a support having thereon a porous spreading layer composed of a particulate structure comprising a plurality of particles being bonded to each other on surface areas of adjacent particles where the adjacent particles are in closest proximity to form a coherent, three dimensional lattice which is essentially non-swellable in an aqueous liquid, and

B. determining total lactate dehydrogenase by detecting the rate of said detectable change.

13. A metod for the determination of total lactate dehydrogenase (LDH) comprising the steps of:

A. in the presence of a substrate for LDH and an indicator material which provides a detectable change in response to the reaction of LDH with said substrate, contacting a dry analytical element with a sample of a liquid suspected of containing LDH, said element comprising a support having thereon

a porous spreading layer composed of a particulate structure comprising a plurality of particles being bonded to each other on surface areas of said particles which are adjacent, where said adjacent particles form a coherent, three dimensional lattice which is essentially non-swellable in an aqueous liquid,

said element further comprising a saturated or unsaturated fluorinated surfactant containing one or more fluorocarbon moieties each having from 4 to 16 carbon atoms and from 8 to 33 fluorine atoms, and each of said moieties having a ratio of fluorine atoms to carbon atoms of about 2;

1, provided that when said surfactant contains more than one such moiety, said moieties have up to two fluorinated or unfluorinated methyl branches, and

B. determining total lactate dehydrogenase by detecting the rate of said detectable change.

14. The method of claim 13 wherein said detectable change is a spectrophotometric change.

15. The method of claim 14 wherein said detectable change is a colorimetric change.

16. The method of claim 15 wherein the rate of disappearance of absorption is detected.

17. A method for the determination of total lactate dehydrogenase (LDH) comprising the steps of:

A. contacting a dry multilayer analytical element with a sample of a liquid suspected of containing LDH to provide a detectable colorimetric change, said element comprising a support having thereon, in order from said support and in fluid contact,

a registration layer comprising an indicator material which provides a detectable spectrophotometric change in response to the reaction of LDH with a substrate for LDH, and

an isotropically porous spreading layer comprising said LDH substrate and composed of a particulate structure comprising;

(i) a plurality of heat-stable, organo-polymeric particles non-swellable in and impermeable to an aqueous liquid, and having a particle size of from about 1 to about 200 μ

m, and

(ii) an adhesive, in an amount of up to about 10 weight percent of said particles, comprising an organic polymer different from that of said particles,

substantially all of said adhesive being concentrated on surface areas of adjacent particles where said adjacent particles are in closest proximity, and bonding said particles into a coherent, three-dimensional lattice which is essentially non-swellable in said liquid,

said element containing, in any of said layers, a saturated or unsaturated fluorinated surfactant containing one or more fluorocarbon moieties each having from 4 to 16 carbon atoms and from 8 to 33 fluorine atoms, and each of said moieties having a ratio of fluorine atoms to carbon atoms of about 2;

1, provided that when said surfactant contains more than one such moiety, said moieties have up to two fluorinated or unfluorinated methyl groups, which surfactant is present in a molar ratio to said substrate of from about 0.01;

1 to about 10;

1, and

B. determining total lactate dehydrogenase by detecting the rate of said colorimetric change.

18. The method of claim 17 wherein said liquid is human serum or whole blood.

19. The method of claim 17 wherein the rate of disappearance of absorption is detected.