Multilayer analytical element

• US 4,066,403 A
• Filed: 05/20/1976
• Issued: 01/03/1978
• Est. Priority Date: 06/20/1975
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Bruschi, Barbara J.
• Primary Examiner(s): Reese, Robert M.
• Application Number:
  US05/688,446
• Time in Patent Office: 593 Days
• Field of Search: 23/253 TP, 23/230 B, 195/103.5, 73/356
• US Class Current: 435/12
• CPC Class Codes: C09B 23/02 the polymethine chain containing an odd number of >CH- or >C[alkyl]- groups
• CPC Class Codes: C09B 23/105 two >CH- groups
• CPC Class Codes: C09B 51/00 Nitro or nitroso dyes
• CPC Class Codes: G01N 31/00 Investigating or analysing non-biological materials by the use of the chemical methods specified in the subgroup; Apparatus specially adapted for such methods
• CPC Class Codes: Y10T 436/171538 Urea or blood urea nitrogen
• CPC Class Codes: Y10T 436/173845 Amine and quaternary ammonium

Abstract

An analytical element for assay of complex fluids, such as blood, the element comprising two reagents and a barrier composition separating the reagents. Such an element is particularly useful in the analysis of blood for urea nitrogen and cholesterol content.

Claims

1. In an integral element useful in the analysis of a fluid sample for a predetermined analyte, the element includinga reagent capable of interacting in the presence of said analyte to produce a decomposition product;

and

2. a reagent capable of interacting with said decomposition product or an intermediate to provide a detectable change;

the improvement comprising

a barrier composition separating substantially all of said said reagent (1) from said reagent (2), said barrier composition being substantially uniformly permeable to said decomposition product and substantially impermeable to interferants.

2. An element as defined in claim 1, wherein said reagents are uniformly distributed in separate layers and said barrier composition is in a layer between said separate layers or in one of said separate layers.

3. An element as defined in claim 2 wherein said barrier composition is substantially impermeable to liquid water and wherein said decomposition product is a vapor.

4. An element as defined in claim 1 wherein said composition includes a member selected from the group consisting of cellulose propionate valerate, cellulose acetate butyrate, cellulose acetate, and poly(methyl methacrylate).

5. In an integral, multilayered element useful in the analysis of a fluid sample for a predetermined analyte, the element including, in superposed relationship:

1. a first layer which includes at least one reagent capable of interacting in the presence of the analyte to produce a decomposition product; and

2. a second layer which includes at least one reagent capable of interacting in the presence of said decomposition product or an intermediate to provide a detectable change;

the improvement comprising

a barrier composition separating substantially all of said reagents in said first and second layers from one another, said composition being substantially uniformly permeable to said decomposition product and substantially impermeable to interferants;

one of said reagents being distributed within said barrier composition.

6. An element as defined in claim 5 wherein said first layer reagent includes at least one enzyme capable of complexing with analyte to produce a decompostion product capable of reacting with said distributed reagent to produce said detectable species;

and further including a non-fibrous isotropically porous spreading layer.

7. An element as defined in claim 6 wherein said enzyme is confined to a discrete layer interposed between said spreading layer and said barrier composition.

8. An element as defined in claim 5 wherein said barrier composition is substantially impermeable to liquid water and wherein said decomposition product is a vapor.

9. An element as defined in claim 8 wherein said composition includes a member selected from the group consisting of cellulose propionate valerate, cellulose acetate butyrate, cellulose acetate, and poly(methyl methacrylate).

10. In an integral, multilayered element useful in the analysis of a fluid sample for a predetermined analyte, the element including, in superposed relationship:

1. a first layer which includes at least one reagent capable of interacting in the presence of the analyte to produce a decomposition product; and

2. a second layer which includes at least one reagent capable of interacting with said decomposition product or an intermediate to provide a detectable change;

the improvement comprising

a barrier composition separating substantially all of said first and second layer reagents from one another, said barrier composition being substantially permeable to said decomposition product and substantially uniformly impermeable to interferants;

said barrier composition being contiguous with and between said two layers.

11. An element as defined in claim 10 wherein said first layer reagent includes at least one enzyme capable of complexing with a predetermined analyte to produce a decomposition product capable of reacting with said second layer reagent to produce said detectable change;

and further including an isotropically porous spreading layer.

12. An element as defined in claim 11 wherein said enzyme is generally confined to a layer distinct from and between said spreading layer and said barrier composition.

13. An element as defined in claim 12 wherein said composition includes a member selected from the group consisting of cellulose propionate valerate, cellulose acetate butyrate, cellulose acetate, and poly(methyl methacrylate).

14. In an integral element useful in the analysis of a fluid sample for urea, the element includingurease;

and

2. a reagent capable of interacting with ammonia to form a detectable product;

the improvement comprising

a barrier composition separating substantially all of said urease from said reagent, said composition being substantially uniformly permeable to ammonia vapor and substantially impermeable to liquid bases.

15. An element as defined in claim 14 wherein said reagent includes a dye precursor which, in the presence of NH₃, produces a dye.

16. An element as defined in claim 15 wherein said dye precursor is a leuco compound which deprotonates in the presence of NH₃.

17. An element as defined in claim 16 wherein said leuco compound is a leuco dye selected from the group consisting of leuco cyanine dyes, nitro-substituted leuco dyes, and leuco phthalein dyes.

18. An element as defined in claim 17 and for the analysis of BUN, wherein said leuco dye is present in an amount sufficient to provide deprotonation in response to all the ammonia transmitted to the leuco dye during the time of the analysis and corresponding to a range of from 0 to about 120 mg of BUN per deciliter of sample.

19. In an integral element useful in the analysis of a fluid sample for urea, the element includingurease;

and

2. a reagent capable of interacting with ammonia to form a detectable product;

the improvement comprising

a barrier composition separating said urease and said reagent, said composition being substantially uniformly permeable to ammonia vapor and substantially impermeable to liquid bases,

said reagent including a 1,2-substituted quinolinium salt which deprotonates in the presence of NH₃ to produce a dye.

20. In an integral element useful in the analysis of a fluid sample for urea, the element includingurease;

and

2. a reagent capable of interacting with ammonia to form a detectable product;

the improvement comprising

a barrier composition separating said urease and said reagent, said composition being substantially uniformly permeable to ammonia vapor and substantially impermeable to liquid bases,

said reagent including a 1,4-substituted quinolinium salt which deprotonates in the presence of NH₃ to produce a dye.

21. In an integral element useful in the analysis of a fluid sample for urea, the element includingurease;

and

2. a reagent capable of interacting with ammonia to form a detectable product;

the improvement comprising

a barrier composition separating said urease and said reagent, said composition being substantially uniformly permeable to ammonia vapor and substantially impermeable to liquid bases,

said reagent including a leuco dye that deprotonates in the presence of NH₃ and has the formula ##STR17## wherein;

a. k represents 0 or 1;

b. m represents 0 or 1;

c. n represents 1 or 2;

d. each L represents a methine group;

e. A represents an electron donating moiety, selected from the group oxygen (--O--), sulfur (--S--), and ##STR18## f. E is an electron-withdrawing group selected from the class consisting of nitro, cyano, ethoxycarbonyl, and halogenated methyl;

g. p represents 0 or 1;

h. R³ represents either an alkyl group having from 1 to 18 carbon atoms and preferably a lower alkyl group having from 1 to 4 carbon atoms, a sulfoalkyl group, a carboxyalkyl group, a sulfatoalkyl group, an alkoxyalkyl group, an acyloxyalkyl group, an alkoxycarbonylalkyl group, a dialkylaminoalkylene group, a cycloaminoalkylene group, an alkenyl group, or an aryl group;

i. X^- is an acid anion;

j. Z³ represents the atoms necessary to complete an aryl ring; and

k. Z⁴ represents the nonmetallic atoms necessary to complete a heterocyclic nucleus.

22. An element as defined in claim 21 wherein said leuco dye is 1-ethyl-4(2,6-dinitrophenyl)methyl quinolinium ethyl sulfonate.

23. An element as defined in claim 21 wherein said leuco dye is 1-methyl-2(2,4-dinitrophenyl)methyl quinolinium perchlorate.

24. An element as defined in claim 21 wherein said leuco dye is 1-ethyl-4(2,4-dinitro-1-naphthyl)methyl quinolinium chloride.

25. An element as defined in claim 21 wherein said leuco dye is 1-propyl-4(2,6-dinitro-4-chlorophenyl)methyl quinolinium ethane sulfonate.

26. In an integral element useful in the analysis of a fluid sample for urea, the element includingurease;

and

2. a reagent capable of interacting with ammonia to form a detectable product;

the improvement comprising

a barrier composition separating said urease and said reagent, said composition being substantially uniformly permeable to ammonia vapor and substantially impermeable to liquid bases,

said reagent including a dye precursor that deprotonates in the presence of NH₃ and has the formula ##STR19## wherein;

n represents a positive integer of 1 to 10;

X^- represents an acid anion;

R¹ and R² are the same or different and each represents either hydrogen, a substituted or unsubstituted alkyl group, an alkoxyalkyl group, a carboxyalkyl group, a sulfoalkyl group, a sulfato alkyl group, an acyloxyalkyl group, an alkoxycarbonylalkyl group, an aralkyl group, an alkenyl group, or a substituted or unsubstituted aryl group; and

Z¹ and Z² are the same or different and each represents the nonmetallic atoms necessary to complete a heterocyclic nucleus.

27. In an integral element useful in the analysis of a fluid sample for urea, the element includingurease;

and

2. a reagent capable of interacting with ammonia to form a detectable product;

the improvement comprising

a barrier composition separating said urease and said reagent, said composition being substantially uniformly permeable to ammonia vapor and substantially impermeable to liquid bases,

said reagent including an ammonia-bleachable dye.

28. An element as defined in claim 27 wherein said dye is a pyrylium dye or styryl dye.

29. An integral multilayered element useful in the analysis of a fluid sample for a predetermined analyte, the element comprising, in superposed relationship,

31. An element as defined in claim 29 wherein said spreading layer includes means for reflecting light.

30. a first layer which includes an enzyme capable of complexing with analyte to produce a decomposition product;

2. a second layer which includes a reagent capable of interacting with said decomposition product or an intermediate to form a detectable change;

3. a barrier composition separating substantially all of said enzyme from said reagent, said composition being substantially uniformly permeable to said decomposition product and substantially impermeable to interferants; and

4. a non-fibrous isotropically porous spreading layer.

32. An element as defined in claim 30 wherein said means includes titanium dioxide.

33. An integral element useful in the analysis of a fluid sample for a predetermined analyte, the element comprisingan enzyme capable of complexing with said analyte to produce a decomposition product;

2. a dye precursor capable of forming a detectable change and having the formula ##STR20## wherein;

n represents a positive integer of 1 to 10;

X^- represents an acid anion;

R¹ and R² are the same or different and each represents either hydrogen, a substituted or unsubstituted alkyl group, an alkoxyalkyl group, a carboxyalkyl group, a sulfoalkyl group, a sulfato alkyl group, an acyloxyalkyl group, an alkoxycarbonylalkyl group, an aralkyl group, an alkenyl group, or a substituted or unsubstituted aryl group; and

Z¹ and Z² are the same or different and each represents the nonmetallic atoms necessary to complete a heterocyclic nucleus; and

34. a barrier composition separating said enzyme and said dye, said composition being substantially uniformly permeable to said decomposition product and substantially impermeable to interferants.

37. An element as defined in claim 34 wherein said barrier composition is substantially impermeable to liquid water and wherein said decomposition product is a vapor.

40. An element as defined in claim 37 wherein said enzyme is confined to a layer separate and distinct from the spreading layer between said spreading layer and said barrier composition.

43. An element as defined in claim 40 wherein said barrier composition is substantially impermeable to liquid water.

44. An element as defined in claim 40 wherein said one reagent includes a leuco compound which deprotonates in the presence of NH₃.

35. A process of analyzing a liquid for the presence of a predetermined analyte, comprising the steps of

a. depositing a sample of the liquid on an element comprising a first reagent capable of producing a radiant energy detectable species and having in association with it a binder, a second reagent capable of producing from the analyte a decomposition product which is interactive with said first reagent;

and a barrier composition separating substantially all of said second reagent from said first reagent, said composition being substantially uniformly permeable to said decomposition product and substantially impermeable to interferants;

whereby the analyte and the second reagent react to produce a decomposition product that selectively permeates through the barrier composition where it reacts with the first reagent; and

b. measuring said detectable change.

36. In an integral, multilayered element useful in the analysis of a fluid sample for a predetermined analyte, the element including, in superposed relationship:

1. a first layer which includes a sample intercept area, and reagent in said area capable of interacting with the analyte to produce a decomposition produt; and

2. a second layer which includes at least one reagent capable of interacting with said decomposition product or an intermediate to form a detectable change;

the improvement comprising

a barier composition separating substantially all of said first and second layer reagent from one another and having a sample intercept area that is at least coextensive with said sample intercept area of said first layer, said composition within its sample intercept area being uniformly permeable to said decomposition product and impermeable to interferants.

39. An element as defined in claim 36 wherein said first layer reagent includes an enzyme capable of complexing with the analyte being tested to effect a decomposition product capable of reacting with said incorporated reagent to produce said detectable species, and further including a non-fibrous isotropically porous spreading layer.

41. An element as defined in claim 36 wherein said barrier composition includes a member selected from the group consisting of cellulose propionate valerate, cellulose acetate butyrate, cellulose acetate, and poly(methyl methacrylate).

38. In an integral, multilayered element useful in the analysis of a fluid sample for a predetermined analyte, the element including, in superposed relationship:

1. a first layer which includes a sample intercept area and a reagent in said area capable of interacting with the analyte to produce a decomposition product; and

2. a second layer which includes a reagent capable of interacting with said decomposition product or an intermediate to form a detectable change;

the improvement comprising

a barrier composition separating substantially all of said first and second layer reagents from one another and having a sample intercept area that is at least coextensive with said sample intercept area of said first layer, said barrier composition within said sample intercept area being uniformly impermeable to interferants;

said barrier composition incorporating one of said reagents within said coextensive sample intercept area.

42. In an integral element useful in the analysis of a fluid sample for urea, the element includinga layer comprising urease in a sample intercept area;

and

2. a reagent capable of interacting with ammonia to form a detectable change;

the improvement comprising a barrier composition separating substantially all of said urease from said reagent and having a sample intercept area that is at least coextensive with the sample intercept area of the layer comprising said urease, said barrier composition within its sample intercept area being uniformly permeable to ammonia vapor and impermeable to liquid base.

45. An element as defined in claim 42 wherein said compound is selected from the group consisting of leuco cyanine dyes, nitro-substituted leuco dyes, and leuco phthalein dyes.

46. In an integral element useful in the analysis of a fluid sample for urea, the element includinga layer comprising urease in a sample intercept area;

and

47. a reagent capable of interacting with ammonia to form a detectable change;

the improvement comprising a barrier composition separating said urease and said reagent and having a sample intercept area that is at least coextensive with the sample intercept area of the layer comprising said urease, said barrier composition within its sample intercept area being uniformly permeable to ammonia vapor and impermeable to liquid bases,

said reagent including a 1,2-substituted quinolinium salt which deprotonates in the presence of NH₃.

48. In an integral element useful in the analysis of a fluid sample for urea, the element includinga layer comprising urease in a sample intercept area;

and

2. a reagent capable of interacting with ammonia to form a detectable change;

the improvement comprising a barrier composition separating said urease and said reagent and having a sample intercept area that is at least coextensive with the sample intercept area of the layer comprising said urease, said barrier composition within its sample intercept area being uniformly permeable to ammonia vapor and impermeable to liquid bases,

said reagent including a 1,4-substituted quinolinium salt which deprotonates in the presence of NH₃.

53. An element as defined in claim 48 wherein said dye is a pyrylium dye or styryl dye.

49. In an integral element useful in the analysis of a fluid sample for urea, the element includinga layer comprising urease in a sample intercept area;

and

2. a reagent capable of interacting with ammonia to form a detectable change;

the improvement comprising a barrier composition separating said urease and said reagent and having a sample intercept area that is at least coextensive with the sample intercept area of the layer comprising said urease, said barrier composition within its sample intercept area being uniformly permeable to ammonia vapor and impermeable to liquid bases,

said reagent including a leuco dye that deprotonates in the presence of NH₃ and has the formula ##STR21## wherein;

a. k represents 0 or 1;

b. m represents 0 or 1;

c. n represents 1 or 2;

d. each L represents a methine group;

e. A represents an electron donating moiety, selected from the group oxygen (--O--), sulfur (--S--), and ##STR22## f. E is an electron-withdrawing group selected from the class consisting of nitro, cyano, ethoxycarbonyl, and halogenated methyl;

g. p represents o or 1;

h. R³ represents either an alkyl group having from 1 to 18 carbon atoms and preferably a lower alkyl group having from 1 to 4 carbon atoms, a sulfoalkyl group, a carboxyalkyl group, a sulfatoalkyl group, an alkoxyalkyl group, an acyloxyalkyl group, an alkoxycarbonyl alkyl group, a dialkylaminoalkylene group, a cycloaminoalkylene group, an alkenyl group, or an aryl group;

i. X^- is an acid anion;

j. Z³ represents the atoms necessary to complete an aryl ring; and

k. Z⁴ represents the nonmetallic atoms necessary to complete a heterocyclic nucleus.

50. In an integral element useful in the analysis of a fluid sample for urea, the element includinga layer comprising urease in a sample intercept area;

and 2. a reagent capable of interacting with ammonia to form a detectable change;

the improvement comprising a barrier composition separating said urease and said reagent and having a sample intercept area that is at least coextensive with the sample intercept area of the layer comprising said urease, said barrier composition within its sample intercept area being uniformly permeable to ammonia vapor and impermeable to liquid bases,

said reagent including a chromogen that deprotonates in the presence of NH₃ and has the formula ##STR23## wherein;

n represents a positive integer of 1 to 10;

X^- represents an acid anion;

R¹ and R² are the same or different and each represents either hydrogen, an alkyl group, an alkoxyalkyl group, a carboxyalkyl group, a sulfoalkyl group, a sulfato alkyl group, an acyloxyalkyl group, an alkoxycarbonylalkyl group, an aralkyl group, an alkenyl group, or an aryl group; and

Z¹ and Z² are the same or different and each represents the nonmetallic atoms necessary to complete a heterocyclic nucleus.

51. In an integral element useful in the analysis of a fluid sample for urea, the element including

52. a layer comprising urease in a sample intercept area;

and

2. a reagent capable of interacting with ammonia to form a detectable change;

the improvement comprising a barrier composition separating said urease and said reagent and having a sample intercept area that is at least coextensive with the sample intercept area of the layer comprising said urease, said barrier composition within its sample intercept area being uniformly permeable to ammonia vapor and impermeable to liquid bases,

said reagent including an ammonia-bleachable dye.

54. An integral element useful in the analysis of a fluid sample for a predetermined analyte, the element comprisinga layer which includes an enzyme in a sample intercept area, capable of reacting with said analyte to produce a decomposition product;

2. a dye precursor having the formula ##STR24## wherein;

n represents a positive integer of 1 to 10;

X^- represents an acid anion;

R¹ and R² are the same or different and each represents either hydrogen, an alkyl group, an alkoxyalkyl group, a carboxyalkyl group, a sulfoalkyl group, a sulfato alkyl group, an acyloxyalkyl group, an alkenyl group, or an aryl group; and

Z¹ and Z² are the same or different and each represents the nonmetallic atoms necessary

to complete a heterocyclic nucleus; and

55. a barrier composition separating said enzyme and said dye and having a sample intercept area that is at least coextensive with said enzyme sample intercept area, said composition within its sample intercept area being uniformly permeable to said decomposition product and impermeable to interferants.