Analytical elements with improved reagent stability

• US 4,283,491 A
• Filed: 08/20/1979
• Issued: 08/11/1981
• Est. Priority Date: 09/06/1977
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Dappen, Glen M.
• Primary Examiner(s): Wiseman, Thomas G.
• Application Number:
  US06/067,843
• Time in Patent Office: 722 Days
• Field of Search: 435/14, 435/28, 435/188, 435/192, 435/805, 435/810, 435/25, 435/10, 422/56, 422/61, 252/408 R
• US Class Current: 435/10
• CPC Class Codes: C12Q 1/28 involving peroxidase
• CPC Class Codes: C12Q 1/54 involving glucose or galactose
• CPC Class Codes: C12Q 1/62 involving uric acid
• 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: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10T 436/108331 Preservative, buffer, anticoagulant or diluent

Abstract

Elements for the analysis of aqueous liquids are described. Deterioration of test reagents, particularly peroxidase, during storage is reduced by inclusion in the elements of a co-polymer derived from hydrophobic, addition-polymerizable monomers.

Claims

1. In an element for the analysis of a predetermined analyte in an aqueous liquid, said element comprising:

(a) a carrier permeable to said aqueous liquid and to said predetermined analyte and,

(b) dispersed in said carrier, reagents capable of interacting with or facilitating interaction with said predetermined analyte or its reaction products to yield a detectable product, one of said reagents comprising peroxidase, the improvement comprising having dispersed in said carrier in addition to said reagents a polymer in an amount sufficient to impart stability to said peroxidase, said amount comprising from about 20 percent to about 50 percent of the total weight of said carrier material plus said polymer, said polymer comprising;

(1) from about 80 to about 98 percent by weight of recurring units derived from one or more hydrophobic, addition-polymerizable monomers selected from the group consisting of an alkyl acrylate, an alkyl methacrylate, styrene and a substituted styrene,

(2) from about 1 to about 20 percent by weight of recurring units derived from one or more anionic monomers selected from the group consisting of an acrylic acid, an acrylic acid salt, a methacrylic acid, a methacrylic acid salt, a sulfonic acid, a sulfonic acid salt, a sulfonate, a sulfate, a phosphate and a phosphorate, and

(3) from 0 to about 15 percent by weight of recurring units derived from one or more crosslinkable, active methylene group-containing monomers.

2. An element as described in claim 1 wherein said reagents further comprise an enzyme capable of catalyzing the oxidation of said predetermined analyte to a peroxide and an indicator composition oxidizable by said peroxide in the presence of peroxidase to yield a detectable product.

3. An element as described in claim 1 wherein said reagents further comprise a compound for adjusting pH.

4. An element as described in claim 1 wherein said carrier comprises a hydrophilic colloid selected from the group consisting of gelatin, a gelatin derivative, a cellulose derivative, a polysaccharide, an acrylamide, poly(vinyl alcohol), and poly(vinyl pyrrolidone).

5. An element as described in claim 1 which comprises more than one layer, said carrier comprising one of said layers.

6. An element as described in claim 1 wherein said carrier comprises a layer coated on a support material.

7. An element as described in claim 1 wherein a subbing layer is coated on said carrier and a spreading-reflecting layer is coated on the subbing layer.

8. In an element for the analysis of a predetermined analyte in an aqueous liquid, said element comprising:

(a) a carrier permeable to said aqueous liquid and to said predetermined analyte and

(b) dispersed in said carrier, reagents capable of interacting with or facilitating interaction with said predetermined analyte or its reaction products to yield a detectable product, one of said reagents comprising peroxidase,

the improvement comprising having dispersed in said carrier in addition to said reagents a polymer comprising;

(1) from about 80 to about 98 percent by weight of recurring units derived from one or more hydrophobic, addition-polymerizable monomers selected from the group consisting of an alkyl acrylate, an alkyl methacrylate, styrene, and a substituted styrene,

(2) from about 1 to about 20 percent by weight of recurring units derived from one or more anionic monomers selected from the group consisting of an acrylic acid, an acrylic acid salt, a methyacrylic acid, a methyacrylic acid salt, a sulfonic acid, a sulfonic acid salt, a sulfonate, a sulfate, a phosphate, and a phosphorate, and

(3) from 0 to about 15 percent by weight of recurring units derived from one or more crosslinkable, active methylene group-containing monomers,

said polymer comprising from about 20 percent to about 50 percent of the total weight of said carrier plus said polymer.

9. An element as described in claim 8 wherein said reagents further comprise an enzyme capable of catalyzing the oxidation of said predetermined analyte to a peroxide and an indicator composition oxidizable by said peroxide in the presence of peroxidase to yield a detectable product.

10. An element as described in claim 8 wherein said reagents further comprise a compound for adjusting pH.

11. An element as described in claim 8 wherein said carrier comprises a hydrophilic colloid selected from the group consisting of gelatin, a gelatin derivative, a cellulose derivative, a polysaccharide, an acrylamide, poly(vinyl alcohol), and poly(vinyl pyrrolidone).

12. An element as described in claim 8 which comprises more than one layer, said carrier comprising one of said layers.

13. An element as described in claim 8 wherein said carrier comprises a layer coated on a support material.

14. An element as described in claim 8 wherein a subbing layer is coated on said carrier and a spreading-reflecting layer is coated on the subbing layer.

17. An element as described in claim 13 wherein said reagents further comprise an enzyme capable of catalyzing the oxidation of said predetermined analyte to a peroxide and an indicator composition oxidizable by said peroxide in the presence of peroxidase to yield a detectable product.

18. An element as described in claim 13 wherein said reagents further comprise a compound for adjusting pH.

19. An element as described in claim 13 wherein said carrier comprises a hydrophilic colloid selected from the group consisting of gelatin, a gelatin derivative, a cellulose derivative, a polysaccharide, an acrylamide, poly(vinyl alcohol), and poly(vinyl pyrrolidone).

20. An element as described in claim 13 which comprises more than one layer, said carrier comprising one of said layers.

21. An element as described in claim 13 wherein said carrier comprises a layer coated on a support material.

22. An element as described in claim 13 wherein a subbing layer is coated on said carrier and a spreading-reflecting layer is coated on the subbing layer.

15. In an element for the analysis of a predetermined analyte in an aqueous liquid, said element comprising:

(a) a carrier permeable to said aqueous liquid and to said predetermined analyte and,

(b) dispersed in said carrier, reagents capable of interacting with or facilitating interaction with said predetermined analyte or its reaction products to yield a detectable product, one of said reagents comprising peroxidase,

the improvement comprising having dispersed in said carrier in addition to said reagents a polymer in an amount sufficient to impart stability to said peroxidase, said amount comprising from about 20 percent to about 50 percent of the total weight of said carrier material plus said polymer, said polymer being selected from the group consisting of;

poly(methyl acrylate-co-3-acryloxyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 85;

10;

5);

poly(n-butyl acrylate-co-3-acryloyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 91.25;

4.75;

4.0); and

poly(n-butyl acrylate-co-2-acrylamido-2-methylpropane sulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 85;

10;

5).

16. In an element for the analysis of a predetermined analyte in an aqueous liquid, said element comprising:

(a) a carrier permeable to said aqueous liquid and to said predetermined analyte and

(b) dispersed in said carrier, reagents capable of interacting with or facilitating interaction with said predetermined analyte or its reaction products to yield a detectable product, one of said reagents comprising peroxidase,

the improvement comprising having dispersed in said carrier in addition to said reagents a polymer selected from the group consisting of

Poly(methyl acrylate-co-3-acryloyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

Poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

Poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio=85;

10;

5);

Poly(n-butyl acrylate-co-3-acryloyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 91.25;

4.75;

4.0); and

Poly(n-butyl acrylate-co-2-acrylamido-2-methylpropane sulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 85;

10;

5),

said polymer comprising from about 20 percent to about 50 percent of the total weight of said carrier plus said polymer.

23. In an element for the analysis of glucose in aqueous liquids, said element comprising:

(a) a carrier permeable to said aqueous liquid and to glucose and,

(b) dispersed in said carrier, reagents comprising glucose oxidase, peroxidase, 7-hydroxy-1-naphthol, 4-aminoantipyrine HCL and a phosphate buffer system (pH

     6),

the improvement comprising having dispersed in said carrier in addition to said reagents a polymer in an amount sufficient to impart stability to said peroxidase, said amount comprising from about 20 percent to about 50 percent of the total weight of said carrier material plus said polymer, said polymer being selected from the group consisting of;

poly(methyl acrylate-co-3-acryloxyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 85;

10;

5);

poly(n-butyl acrylate-co-3-acryloyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 91.25;

4.75;

4.0); and

poly(n-butyl acrylate-co-2-acrylamido-2-methylpropane sulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 85;

10;

5).

24. An element as described in claim 23 wherein said carrier comprises a hydrophilic colloid selected from the group consisting of gelatin, a gelatin derivative, a cellulose derivative, a polysaccharide, an acrylamide, poly(vinyl alcohol), and poly(vinyl pyrrolidone).

25. An element as described in claim 23 wherein said polymer comprises from about 20 percent to about 50 percent of the total weight of said carrier plus said polymer.

26. An element as described in claim 23 wherein the carrier is a layer comprising deionized gelatin coated on a polyethylene terephthalate film support and said carrier is overcoated with a subbing layer and a blushed-polymer, spreading-reflecting layer comprising cellulose acetate and titanium dioxide.

27. In an element for the analysis of uric acid in aqueous liquids, said element comprising:

(a) a carrier permeable to said aqueous liquid and to uric acid and,

(b) dispersed in said carrier, reagents comprising bis(vinylsulfonylmethyl)ether, uricase, peroxidase, a dispersion of 2-(3,5-dimethoxy-4-hydroxyphenyl)-4,5-bis-(4-dimethylaminophenyl)imidazole in 2,4-di-n-pentylphenol, and a borate buffer (pH

     9),

the improvement comprising having dispersed in said carrier in addition to said reagents a polymer in an amount sufficient to impart stability to said peroxidase, said amount comprising from about 20 percent to about 50 percent of the total weight of said carrier material plus said polymer, said polymer being selected from the group consisting of;

poly(methyl acrylate-co-3-acryloxyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 88.75;

4.75;

6.5);

poly(methyl acrylate-co-2-acrylamido-2-methyl.tbd.propanesulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 85;

10;

5);

poly(n-butyl acrylate-co-3-acryloyloxypropane.tbd.sulfonic acid, sodium salt-co-2-acetoacetoxyethyl methacrylate) (weight ratio 91.25;

4.75;

4.0); and

poly(n-butyl acrylate-co-2-acrylamido-2-methylpropane sulfonic acid-co-2-acetoacetoxyethyl methacrylate) (weight ratio 85;

10;

5).

28. An element as described in claim 27 wherein said carrier comprises a hydrophilic colloid selected from the group consisting of gelatin, a gelatin derivative, a cellulose derivative, a polysaccharide, an acrylamide, poly(vinyl alcohol), and poly(vinyl pyrrolidone).

29. An element as described in claim 27 wherein said polymer comprises from about 20 percent to about 50 percent of the total weight of said carrier plus said polymer.

30. An element as described in claim 27 wherein the carrier is a layer comprising deionized gelatin coated on a polyethylene terephthalate film support and said carrier is overcoated with a layer comprising deionized gelatin, bis(vinylsulfonylmethyl) ether, borate buffer (pH9), a subbing layer comprising poly(n-isopropyl acrylamide) and a blushed-polymer, spreading-reflecting layer comprising cellulose acetate and titanium dioxide.