Element for analysis of liquids

• US 4,144,306 A
• Filed: 02/13/1978
• Issued: 03/13/1979
• Est. Priority Date: 01/14/1977
• Status: Expired due to Term

• Current Assignee: Clinical Diagnostic Systems Inc.
• Original Assignee: Eastman Kodak Company
• Inventors: Figueras, John
• Primary Examiner(s): Wolk, Morris O.
• Assistant Examiner(s): Marcus, Michael S.
• Application Number:
  US05/877,193
• Time in Patent Office: 393 Days
• Field of Search: 23/253 TP, 23/231, 195/103.5 R, 195/103.5 S, 195/103.5 A
• US Class Current: 422/423
• CPC Class Codes: C12Q 1/40 involving amylase
• CPC Class Codes: G01N 33/04 Dairy products
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: Y10T 436/144444 Glucose

Abstract

A multilayer analytical element for the analysis of liquids, particularly small samples of biological liquids, having at least two layers including a reagent layer and a registration layer. The reagent layer contains an interactive composition including a nondiffusible material having a preformed detectable moiety, such composition being interactive in the presence of liquid containing an analyte of choice to provide a diffusible product comprising the preformed detectable moiety. The registration layer receives the diffusible product released from the reagent layer. The layers present in the analytical element are composed such that the detectable moiety released from the reagent layer or that remaining unreleased in the reagent layer can be selectively detected in the element.

In one embodiment, there is disclosed an integral element which can include a support, preferably radiation-transmissive, on which a registration layer and a reagent layer are carried. Optionally, a spreading or analyte metering layer can be provided in the element adjacent the reagent layer to facilitate delivery of a uniform concentration of analyte to the reagent layer. In a preferred embodiment of this integral element, a radiation-blocking layer, permeable to the diffusible product, can be provided in the element intervening the reagent layer and the registration layer. The radiation-blocking layer, such as an opaque reflecting layer, can enhance the detection of the preformed detectable moiety in the registration layer or in the reagent layer by reflection densitometry or other appropriate radiometric technique.

The multilayer element of the invention is particularly useful, for example, in the assay of amylase as well as a variety of other analytes.

Claims

1. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising a reagent layer and a registration layer,

(a) said reagent layer comprising a composition including a non-diffusible material comprising a preformed, detectable moiety said composition being interactive in the presence of liquid containing said analyte to provide a diffusible species comprising the preformed, detectable moiety and

(b) said registration layer adapted to receive said diffusible species,

wherein layers within the element are composed such that said preformed, detectable moiety released from the reagent layer can be detected selectively within the element.

2. An analytical element as described in claim 1 wherein the registration layer is radiation-transparent.

3. An analytical element as described in claim 1 wherein the registration layer is at least as swellable in liquid under analysis as is the reagent layer.

4. An analytical element as described in claim 1 wherein the preformed, detectable moiety comprises a radiation emissive moiety or a colorimetrically detectable moiety.

5. An analytical element as described in claim 1 wherein said element comprises a radiation-transmissive support bearing said reagent layer and said registration layer.

6. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising a support having superposed thereon a reagent layer and a registration layer,

(a) said reagent layer comprising a composition including a non-diffusible material comprising a preformed, detectable moiety, said composition being interactive in the presence of liquid containing said analyte to provide a diffusible species comprising the preformed, detectable moeity, and

(b) said registration layer adapted to receive said diffusible species,

wherein the registration layer is interposed between the reagent layer and the support and the reagent layer is removable from the registration layer, and wherein layers within the element are composed such that said preformed, detectable moiety can be selectively detected within the registration layer.

7. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising a radiation-transmissive support having superposed thereon a reagent layer and a registration layer,

(a) said reagent layer comprisingd a composition including a non-diffusible material comprising a preformed, detectable moiety, said composition being interactive in the presence of liquid containing said analyte to provide a diffusible species comprising the preformed detectable moiety, and

(b) said registration layer adapted to receive said diffusible species,

wherein the registration layer is interposed between the support and the reagent layer and wherein the layers of the element are composed such that said preformed, detectable moiety can be selectively detected within the registration layer.

8. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising a radiation-transmissive support having superposed thereon a reagent layer, a radiation-blocking layer, and a registration layer,

(a) said reagent layer comprising a composition including a non-diffusible material comprising a preformed, detectable moiety, said composition being interactive in the presence of liquid containing analyte to provide a diffusible species comprising the detectable moiety, and

(b) said radiation-blocking layer, permeable to the diffusible species, comprising an opacifying agent, and

(c) said registration layer adapted to receive said diffusible species,

wherein the registration layer is the closest to the support of said layers and the radiation-blocking layer is interposed between the registration layer and the reagent layer, and wherein said preformed, detectable moiety can be selectively detected within the registration layer.

9. An analytical element as described in claim 8 wherein the opacifying agent comprises a pigment.

10. An analytical element as described in claim 8 wherein the opacifying agent comprises a reflective pigment.

11. An integral multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising

(a) an isotropically porous spreading layer comprising a non-fibrous material,

(b) a reagent layer comprising a composition comprising a non-diffusable material including a preformed, detectable moiety, said composition being interactive in the presence of liquid containing said analyte to provide a diffusible product comprising the detectable moiety, and

(c) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein the reagent layer is interposed between the spreading layer and the registration layer, and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

13. An analytical element as described in claim 11 wherein said element comprises a radiation-transmissive support, the registration layer being adjacent the support.

12. An integral multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising

(a) an isotropically porous spreading layer comprising a non-fibrous material,

(b) a reagent layer comprising a composition comprising a non-diffusible material including a performed, detectable moiety, said composition being interactive in the presence of liquid containing said analyte to provide a diffusible product comprising the detectable moiety, and

(c) a radiation-blocking layer, permeable to the diffusible product, comprising an opacifying agent,

(d) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein the reagent layer is interposed between the spreading layer and the radiation-blocking layer, and the radiation-blocking layer is interposed between the reagent layer and the registration layer, and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

14. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising

(a) a porous reagent layer comprising a composition comprising a non-diffusable material including a preformed, detectable moiety, said composition being interactive in the presence of liquid containing said analyte to provide a diffusible product comprising the preformed, detectable moiety, and

(b) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein the preformed, detectable moiety can be detected selectively within the registration layer.

15. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising,

(a) a porous, reagent layer comprising a composition comprising a non-diffusible material including a preformed, detectable moiety, said composition being interactive in the presence of liquid containing said analyte to provide a diffusible product comprising the preformed, detectable moiety,

(b) a radiation-blocking layer, permeable to the diffusible product, comprising an opacifying agent, and

(c) a registration layer, permeable to the diffusible product, comprising a water-swellable, radiation-transmissive material.

wherein the radiation-blocking layer is interposed between the reagent layer and the registration layer, and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

16. An analytical element as described in claim 15 wherein said element comprises a radiation-transmissive support, the registration layer being interposed between the support and the radiation-blocking layer.

17. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising a radiation-transmissive support having thereon,

(a) a porous reagent layer comprising a composition comprising a non-diffusible material including a preformed, detectable moiety, said composition being interactive in the presence of liquid containing said analyte to provide a diffusible product comprising the preformed, detectable moiety,

(b) a radiation-blocking layer, substantially impermeable to said non-diffusible material and permeable to said diffusible product, comprising an opacifying agent, and

(c) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein the registration layer is adjacent the support and the radiation-blocking layer is interposed between the registration layer and the reagent layer, and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

18. A multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising,

(a) an isotropically porous reagent layer comprising a composition comprising a non-diffusible material including a preformed, detectable moiety, said composition being interactive in the presence of a liquid containing said analyte to provide a diffusible product comprising the preformed, detectable moiety, and

(b) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein the preformed, detectable moiety can be detected selectively within the registration layer.

19. An integral multilayer analytical element for analysis of liquid containing a predetermined analyte, the element comprising a radiation-transmissive support having thereon,

(a) an isotropically porous, non-fibrous reagent layer comprising a composition comprising a non-diffusible material including a preformed, detectable moiety, said composition being interactive in the presence of a liquid containing said analyte to provide a diffusible product comprising the preformed, detectable moiety,

(b) a radiation-blocking layer, permeable to the diffusible product, comprising an opacifying agent, and

(c) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein the registration layer is adjacent the support and the radiation-blocking layer is interposed between the reagent layer and the registration layer, and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

20. An analytical element as described in claim 19 wherein the opacifying agent comprises a pigment.

21. An analytical element as described in claim 19 wherein the opacifying agent comprises a relfective pigment.

22. An analytical element as described in claim 19 wherein said preformed detectable moiety comprises a radiation-emissive moiety or a colorimetrically detectable moiety.

23. An analytical element as described in claim 19 wherein the registration layer comprises a hydrophilic colloid.

24. An integral multilayer element for analysis of liquid containing a predetermined analyte, the element comprising a radiation-transmissive support having thereon,

(a) an isotropically porous, non-fibrous reagent layer comprising a porous polymeric composition or particulate matter and a composition comprising a non-diffusible material including a preformed, detectable moiety, the latter said composition being interactive in the presence of liquid containing said analyte, to provide a diffusible product comprising the preformed, detectable moiety, and

(b) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein the registration layer is interposed between the support and the reagent layer and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

25. An integral analytical element as described in claim 24 wherein said preformed detectable moiety comprises a radiation-emissive moiety or a colorimetrically detectable moiety.

26. An integral analytical element as described in claim 24 wherein the reagent layer includes a surfactant.

27. An integral analytical element as described in claim 24 wherein the reagent layer comprises particulate matter.

28. An integral analytical element as described in claim 24 wherein the reagent layer comprises particulate material selected from the group consisting of a microcrystalline colloid derived from a polymer, resinous beads and glass beads;

and wherein the registration layer comprises a hydrophilic colloid.

29. An element as described in claim 24 wherein the reagent layer comprises microcrystalline cellulose particles and the registration layer comprises a hydrophilic colloid.

30. An element as described in claim 24 wherein the registration layer comprises a mordant for the diffusible product.

31. An integral analytical element as described in claim 24 wherein said element comprises a non-fibrous radiation-blocking layer, impermeable to said non-diffusible material and permeable to the diffusible product, comprising an opacifying agent, said radiation-blocking layer being interposed between the reagent layer and the registration layer.

32. An integral element as described in claim 31 wherein the opacifying agent comprises a pigment.

33. A multilayer analytical element for detecting amylase in liquids, the element comprising,

(a) a reagent layer comprising a composition comprising a non-diffusible polysaccharide material including a preformed, detectable moiety, said composition being interactive in the presence of a liquid containing amylase to provide a diffusible moiety comprising the preformed, detectable moiety,

(b) a registration layer, permeable to the diffusible product, comprising a radiation-transmissive material,

wherein layers within the element are composed such that said preformed, detectable moiety released from the reagent layer can be detected selectively within the element.

34. A multilayer analytical element as described in claim 33 wherein said element includes a radiation-blocking layer, permeable to the diffusible product, comprising an opacifying agent, said radiation-blocking layer being interposed between the reagent layer and the registration layer.

35. A multilayer analytical element as described in claim 33 wherein the non-diffusible polysaccharide material comprises starch having said detectable moiety chemically bonded thereto.

36. A multilayer integral analytical element as described in claim 33 wherein the detectable moiety comprises a colorimetrically detectable dye moiety or a fluorescent moiety.

37. A multilayer element for detecting amylase in liquids, the element comprising a radiation-transmissive support having thereon,

(a) an isotropically porous reagent layer comprising a porous polymeric composition or particulate matter and a composition comprising a non-diffusible starch material including a preformed, detectable moiety, said composition being interactive in the presence of liquid containing amylase to provide a diffusible product comprising the preformed, detectable moiety,

(b) a non-fibrous, radiation-blocking layer, permeable to the diffusible product, comprising an opacifying agent, and

(c) a registration layer, permeable to the diffusible product, comprising a water-swellable, radiation-transmissive material,

wherein the registration layer is interposed between the support and the radiation-blocking layer and the radiation-blocking layer is interposed between the reagent layer and the registration layer, and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

38. An analytical element as described in claim 37 wherein the isotropically porous reagent layer comprises the interactive composition and particulate matter comprising a microcrystalline colloid derived from a polymer.

39. An analytical element as described in claim 37 wherein the registration layer comprises a hydrophilic colloid selected from the group consisting of an acrylamide polymer, gelatin, agarose, dextran and a water-swellable polyvinyl compound.

40. An analytical element as described in claim 37 wherein the opacifying agent comprises a pigment selected from the group consisting of carbon, titanium dioxide, zinc oxide, lead oxide, and barium sulfate.

41. An integral multilayer analytical element for detecting amylase in liquids, the element comprising a radiation-transmissive support having thereon,

(a) an isotropically porous reagent layer comprising particulate matter, a surfactant and a non-diffusible composition comprising starch having chemically bonded thereto a preformed, detectable moiety that is a colorimetrically detectable dye moiety or a fluorescent moiety, said starch being interactive in the presence of liquid containing amylase to provide a diffusible product comprising the preformed, detectable moiety,

(b) a non-fibrous radiation-blocking layer, impermeable to said non-diffusible composition and permeable to the diffusible product, comprising a pigment, and

(c) a registration layer, permeable to the diffusible product, comprising a hydrophilic colloid,

wherein the radiation-blocking layer is interposed between the registration layer and the isotropically porous reagent layer, wherein the registration layer is interposed between the support and the radiation-blocking layer, and wherein the preformed, detectable moiety can be detected selectively within the registration layer.

42. An integral analytical element as described in claim 41 wherein the registration layer comprises a mordant for the diffusible product.

43. An integral analytical element as described in claim 41 wherein the registration layer comprises a hydrophilic colloid selected from the group consisting of an acrylamide polymer, gelatin, agarose, dextran and a water-swellable polyvinyl compound.

44. An integral multilayer element for detecting amylase in liquids, the element comprising a radiation-transmissive support having thereon,

(a) an isotropically porous reagent layer comprising particulate matter, a surfactant and non-diffusible starch having a dye moiety chemically bonded thereto, said starch being interactive in the presence of liquids containing amylase to provide a diffusible product comprising the dye moiety,

(b) a non-fibrous, radiation-blocking layer, permeable to the diffusible product, comprising titanium dioxide and gelatin, and

(c) a non-fibrous, radiation-transmissive registration layer comprising gelatin and poly-(styrene-co-N-vinylbenzyl-N,N-dimethylbenzyl-ammonium chloride-co-divinylbenzene),

wherein the registration layer is adjacent the support and the radiation-blocking layer is interposed between the reagent layer and the registration layer, and wherein the dye moiety can be detected selectively within the registration layer.

45. A method for detecting amylase in liquids, the method comprising the steps of

(a) contacting a sample of liquid under analysis and a reagent layer of a multilayer analytical element comprising (i) said reagent layer and (ii) a registration layer, said reagent layer comprising a composition comprising a non-diffusible polysaccharide material including a preformed, detectable moiety, said composition being interactive in the presence of liquid containing amylase to provide a diffusible product comprising the preformed, detectable moiety, and said registration layer, permeable to the diffusible product, comprising a radiaton-transmissive material, wherein layers within the element are composed such that the preformed, detectable moiety released from the reagent layer can be detected selectively within the element, and

(b) detecting, after a predetermined time, said preformed, detectable moiety.