JPH01265159A - Integral type multilayered analysis element - Google Patents

Abstract

PURPOSE:To obtain the integral type multilayered analysis element which can develop a spotted sample uniformly to a circular shape even if the sample is a whole blood sample by using a woven fabric, at least either of the weft and warp of which consists of plural pieces of yarn, as a development layer. CONSTITUTION:The integral type multilayered analysis element constituted by laminating at least one reagent layer contg. at least one reagent in a binder and the liquid sample development layer consisting of the woven fabric is characterized by using plural pieces of the yarn to constitute at least either of the weft or warp of the woven fabric which constitutes the liquid sample development layer of the above-mentioned element. For example, the aq. liquid sample drops of whole blood, blood plasma, serum, urine or the like is spotted to the development layer and is incubated. The coloration or discoloration in the element is then measured by opposite photometry from the light transmittable base side by using visible light or the UV light of the absorption max. wavelength or the wavelength near the same. The content of the analyte in the liquid sample is determined by the principle of a colorimetric measurement method using a previously formed calibration curve.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Application Field] The present invention relates to an improved dry integrated multilayer analytical element for analyzing specific components in liquid samples, and more particularly for the analysis of samples in aqueous liquids, particularly for the analysis of samples in aqueous liquids. The present invention relates to an integrated multilayer analysis element useful for clinical tests using body fluids (e.g., blood (whole blood, plasma, serum), lymph fluid, saliva, urine, etc.) as samples.

[Conventional technology]

One form of conventional dry analysis element is a water-absorbing reagent layer containing a color reaction reagent and a hydrophilic polymer binder on a transparent support, and a porous development layer (hereinafter sometimes referred to as development layer) as the outermost layer. Many integrated multilayer analysis elements (hereinafter sometimes referred to as multilayer analysis elements) have been proposed. The spreading layer of a multilayer analytical element is used to store an aqueous liquid sample (e.g., blood (whole blood, plasma, serum), lymph, saliva, spinal fluid, vaginal liquid,
(urine; drinking water, alcoholic beverages; river water; industrial wastewater, etc.) is spread horizontally without substantially unevenly distributing the components contained in the aqueous liquid sample, and the hydrophilic polymer is added at a rate of approximately constant volume per unit area. This is a layer that functions to supply (metering function) to a water-absorbing reagent layer or water-absorbing layer containing water. Among the porous expansion layers, Japanese Patent Application Laid-Open No. 49-53888
3-21677), 'Membrane filter (brushed polymer) disclosed in U.S. Patent No. 3,992.158, etc.
Non-fibrous isotropic microporous media layer represented by
There are various types of non-fibrous porous spread layers, such as the continuous void-containing three-dimensional lattice granular structure layer, which is made up of polymer microbeads bonded in point contact with a water-insoluble adhesive, as disclosed in No. 5-90859. However, when whole blood is used as a sample, solid components such as red blood cells in the blood clog the micropores, impairing the spreading action, and the polymer component protein Even when using plasma or serum containing a high concentration of , there is a problem in that the micropores are clogged by the polymeric components, impairing the spreading effect. On the other hand, JP-A-55-164356 and JP-A-57-663
Porous spreading layer (woven spreading layer) made of the woven fabric disclosed in No. 59 etc.
When blood is used as an aqueous liquid sample, whole blood, plasma,
It is a porous spreading layer that can spread any serum well and is excellent in various respects such as ease of manufacturing an integrated multilayer analytical element and durability of the element.

[Problem to be solved by the invention]

As mentioned above, the spreading layer made of textiles has excellent performance, but due to the non-uniformity of vertical and horizontal spreading, the unfolded shape does not become a neat circle but an ellipse, so it is particularly limited. In analysis using area, it has become clear that the long axis direction causes non-uniformity in color development due to the edge effect of the analysis element, and ultimately becomes a factor that causes variations in the analysis results.

An object of the present invention is to provide an integrated multilayer analytical element with good reproducibility by suppressing the occurrence of variations due to excessive expansion in one direction when a liquid sample is expanded into a circular shape and formed into an analytical element. It is in.

Another object of the present invention is to provide an integrated multilayer analysis element that can perform accurate measurements even when using whole blood samples.

Another object of the present invention is to provide an integrated multilayer analysis element that allows easy correlation between measured values of whole blood and measured values of plasma or serum, or mutual conversion of constant values on both sides when blood is used as an aqueous liquid sample. It's about doing.

Other objects of the invention will become apparent from the following description and examples.

[Means to solve the problem]

The integrated multilayer analysis element of the present invention is characterized by the use of a woven fabric in which at least one of the weft and warp consists of a plurality of yarns as the porous spreading layer. That is, the present invention provides an integrated multilayer analysis element in which at least one reagent layer containing at least one reagent in a binder and a liquid sample spreading layer made of fabric are laminated in this order, in which the liquid sample spreading layer is laminated in this order. This invention relates to an integrated multilayer analytical element with improved liquid sample deployability, characterized in that the woven fabric consists of a plurality of yarns (one of which is a small number of weft yarns and warp yarns).

Representative embodiments of the integrated multilayer analytical element of the present invention are as follows.

(1) An integrated multilayer analytical element in which a water absorption layer containing a hydrophilic polymer binder and a porous spreading layer are laminated in this order on a light-transmitting water-impermeable support, wherein the spreading layer is An integrated multilayer analytical element (2) in which at least one of the weft and warp is a woven fabric consisting of a plurality of yarns, which reacts with components in a liquid sample on a light-transparent water-impermeable support to produce a detectable change. An integrated multilayer comprising at least one water-absorbing or water-permeable reagent layer containing at least two reagents and a porous spreading layer in this order, the spreading layer being a woven fabric consisting of a plurality of yarns of at least one of weft and warp. Analytical Element (3) A porous layer comprising a water-absorbing layer comprising a hydrophilic polymeric binder on a light-transparent water-impermeable support and at least one reagent that reacts with a component in the liquid sample to produce a detectable change. This is an integrated multilayer analysis element in which the development layers are laminated and integrated in this order, and the development layer is a woven fabric in which at least one of the weft and the warp is composed of a plurality of yarns.

As the light-transparent water-impermeable support of the integrated multilayer analytical element of the present invention, a water-impermeable transparent support such as that used in the multilayer analytical element disclosed in the above-mentioned patent specifications etc. may be used. I can do it. Examples of such materials include polyethylene terecrate, polycarbonate of bisphenol A, polystyrene, and cellulose esters (e.g., cellulose diacetate, cellulose triacetate, cellulose acetate propionate, etc.) with a thickness of approximately 5 mm.
0 μm to about 1 mm, preferably about 80 μm to about 30 μm
It is possible to use a transparent support in the range of 0 pI11.

If necessary, a known undercoat layer or adhesive layer can be provided on the surface of the support according to the above-mentioned patent specifications, etc., to strengthen the adhesion with the water absorption layer, reagent layer, etc. provided on the support. .

The integrated multilayer analytical element of the present invention may not have a transparent support if it is strong enough to maintain its shape without the transparent support. However, in many cases, it is preferred that the integrated multilayer analytical element of the present invention has a support and that the support and the layer thereon are substantially uniformly adhered.

The water-absorbing layer is a layer mainly composed of a hydrophilic polymer that swells when it absorbs water, and is a layer that can absorb water that has reached or permeated the interface of the layer. The hydrophilic polymer that can be used in the water-absorbing layer is a natural or synthetic hydrophilic polymer having a swelling rate at 30°C in the range of about 150% to about 2000%, preferably about 250% to about 1500%. be. As an example, JP-A-59-171
Gelatin (e.g., acid-treated gelatin, deionized gelatin, etc.), gelatin derivatives (e.g., phthalated gelatin, hydroxyacrylate grafted gelatin, etc.), agarose, pullulan derivatives, polyacrylamide, Examples include polypyryl alcohol and polyvinylpyrrolidone. The dry thickness of the water absorption layer is about 1/M to about 100 μm, preferably about 3 μl to about 50 μl, particularly preferably about 5 μm to about 3 μl.
It is preferably in the range of 0 pm and substantially transparent. The water-absorbing layer may contain a known mordant, basic polymer, acidic polymer, etc.

A water-absorbing or water-permeable reagent layer (hereinafter sometimes referred to as a reagent layer) is a substance containing at least one reagent and a hydrophilic polymeric binder that reacts with the component to be measured in a liquid sample to produce a detectable change. A layer that is either non-porous and water-absorbing or microporous and water-permeable. Detectable changes mainly mean changes that can be detected by optical measurement methods, such as color changes, color development (color development), fluorescence generation,
These include changes in absorption wavelength in the ultraviolet region and occurrence of turbidity.

The reagents contained in the reagent layer are determined by the component to be analyzed in the liquid sample and the chemical reaction selected to analyze this component, and the selected chemical reactions can be contained in two or more separate layers. Examples of reagents to be contained in the reagent layer include reagent compositions containing enzymes disclosed in the above-mentioned patent specifications and other known analytical reagent systems or clinical biochemical diagnostic reagent systems.

The hydrophilic polymer used in the substantially non-porous, water-absorbing reagent layer acts as a medium to substantially uniformly dissolve or disperse the reagent or reagent composition, and also absorbs and absorbs water in the liquid sample. It also has at least the function of allowing the test component to reach the reagent layer. The hydrophilic polymer can be appropriately selected from the same polymers as the water-absorbing hydrophilic polymer used in the water-absorbing layer.

The dry thickness of the substantially non-porous reagent layer is from about 3 μm to about 5 μm.
01! m, preferably in the range of about 5 μm to about 30 μm, and preferably the reagent layer is substantially transparent.

The microporous and water-permeable reagent layer may be made of the same fabric as the woven fabric spreading layer described later, the woven fabric described in JP-A-55-164356, etc., or the organic polymer fiber pulp-containing paper described in JP-A-57-148250. Fibrous porous layer such as a porous layer formed by coating a dispersion of fibers and hydrophilic polymers described in papermaking, JP-A-57-125847, etc.; JP-A-53-216;
77, U.S. Patent No. 3,992.158, etc. Membrane filter layer (brushish polymer layer), isotropic continuous microvoid containing fine particles such as polymer microbeads bonded in point contact with a hydrophilic polymer binder A layer in which a reagent composition is contained in a microporous layer such as a non-fibrous porous layer (three-dimensional lattice-like granular structure layer), or a layer composed of solid fine particles and a hydrophilic polymer binder. This is a layer in which a microporous structure contains a reagent composition. The dry layer thickness of the microporous reagent layer ranges from approximately 7 μm to approximately 250 tp.
vh, preferably in the range of about 10 μm to about 250 μm. When a woven fabric spreading layer is provided on the microporous reagent layer, the space between the two layers is as follows.
It is preferable to use porous adhesive as described in 38756 and the like.

The reagent layer can contain a known pH buffer composition, a polymer pH buffer, a basic polymer, an acidic polymer, a polymer mordant, and the like.

Both the water-absorbing layer and the substantially non-porous reagent layer can be provided on the above-described light-transparent water-impermeable support by a known coating method. Microporous reagent layer is disclosed in JP-A-59-1
20957, JP-A-59-145965, and the like. If necessary, the surface of the support may be subjected to a known physicochemical activation treatment or chemical activation treatment, or a transparent undercoat layer (e.g., gelatin undercoat layer) may be provided to form a water-absorbing layer or support. The adhesive force between the reagent layer and the reagent layer can be strengthened. A water-absorbing layer can be provided between the support and the reagent layer, and when the reagent layer is a microporous reagent layer, it is preferable to provide the water-absorbing layer between the reagent layer and the support.

Both the water-absorbing layer and the substantially non-porous reagent layer can be provided on the above-described light-transparent water-impermeable support by a known coating method. Microporous reagent layer is disclosed in JP-A-59-1
20957, JP-A-59-145965, and the like. If necessary, the surface of the support may be subjected to a known physicochemical activation treatment or chemical activation treatment, or a transparent undercoat layer (e.g., gelatin undercoat layer) may be provided to form a water-absorbing layer or support. A water-absorbing layer can be provided between the reagent layer and the reagent layer, and when the reagent layer is a microporous reagent layer, it is preferable to provide a water-absorbing layer between the reagent layer and the support.

A light shielding layer can be provided on the reagent layer or the water absorption layer. The light shielding layer is a water permeable layer in which fine particles or fine powder (hereinafter simply referred to as fine particles) having light shielding properties or both light shielding properties and light reflecting properties are dispersed and held in a small amount of a hydrophilic polymer binder that has the ability to form a film. is a permeable or water-permeable layer. The light shielding layer detects the color of the aqueous liquid sample that is spotted and supplied to the porous development layer, which will be described later, when measuring detectable changes (color change, color development, etc.) in the reagent layer or water absorption layer from the light-transmitting support side. In particular, it blocks the red color of hemoglobin contained in a whole blood sample, and also functions as a light reflecting layer or a background layer.

Examples of fine particles that have both light-shielding and light-reflecting properties include titanium dioxide fine particles (rutile type, anatase type, or pulcite type microcrystalline particles with a diameter of 0.1 to about 1.2 m), barium sulfate. Examples of light-shielding particles include carbon black, gas black, and carbon microbeads, and among these, titanium dioxide particles and barium sulfate particles are preferred. Hydrophilic polymer binders with film-forming ability include the same hydrophilic polymers used for the water absorption layer, as well as weakly hydrophilic regenerated cellulose, cellulose acetate, etc. Among these, gelatin, gelatin derivatives, polyacrylamide, etc. is preferred. Gelatin and gelatin derivatives can be used in combination with a known hardening agent (crosslinking agent).

The light-shielding layer can be provided by applying an aqueous dispersion of light-shielding fine particles and a hydrophilic polymer onto the reagent layer or water-absorbing layer by a known method and drying it. The dry volume ratio of the light-shielding fine particles to the hydrophilic polymer binder is approximately 10 parts of the light-shielding fine particles to 2.0 parts of the polymer binder.
When the light-shielding fine particles are titanium dioxide fine particles, the weight ratio of the polymer binder is about 0.5 to about 7.5, preferably about 3.0 to about 6.5. 6 to about 1.8, preferably about 0.8
to about 1.5. The thickness of the dry layer of the light shielding layer is about 3 μm to about 3 μm, preferably about 5 pm to about 20 μm.
The pI is in the range of 11.

An adhesive layer can be provided on the reagent layer, water absorption layer, or light shielding layer in order to adhere and laminate a porous development layer, which will be described later. An adhesive layer is essential when a porous spreading layer is provided on a light shielding layer. The adhesive layer consists primarily of a hydrophilic polymer that is capable of adhering and integrating the porous spreading layer when wetted with water or swollen with water. Hydrophilic polymers that can be used in the adhesive layer include the same hydrophilic polymers that can be used in the water-absorbing layer, and among these, gelatin, gelatin derivatives, polyacrylamide, and the like are preferred. The dry layer thickness of the adhesive layer is about 0.5 to about 20 μm.
, preferably in the range of about 1 μm to about 10 μm. The adhesive layer can contain a surfactant. The surfactant is preferably a nonionic surfactant, particularly a nonionic surfactant having a chain structure of 8 to 15 oxyethylene or oxypropylene groups.

The adhesive layer can be provided by applying an aqueous solution containing a hydrophilic polymer and optionally added surfactant etc. onto the reagent layer, water absorbing layer or light shielding layer by a known method.

Directly or via an adhesive layer on the water absorption layer or reagent layer,
Alternatively, a porous spreading layer (fabric spreading layer) made of woven fabric is provided on the light shielding layer (light reflection N) via an adhesive layer.

The porous spreading layer spreads an aqueous liquid sample spotted on its upper surface (the surface far from the yz-light support) in the lateral direction without substantially unevenly distributing the components contained therein. It is a layer that functions to supply (spreading function or metering function) to a water-absorbing layer containing a hydrophilic polymer binder or a water-absorbing or water-permeable reagent layer at a rate of approximately constant volume per unit area. Furthermore, the porous spreading layer is a layer that also functions to filter out non-dissolved substances (eg, red blood cells in a whole blood sample, etc.) that interfere with analysis in an aqueous liquid sample.

As the woven fabric that can be used for the porous spreading layer, a plain woven fabric in which at least one of the weft and warp yarns is composed of a plurality of yarns is used.

Preferred specific examples of plain woven fabrics include plain woven fabrics woven with two wefts and one warp, plain woven fabrics woven with one weft and two warps, and plain woven fabrics woven with two wefts and two warps. be.

The threads that make up the textile fabric include natural fiber threads such as cotton, navy blue and wool, regenerated cellulose such as viscose rayon and cupro, semi-synthetic organic polymers such as cellulose diacetate and cellulose triacetate, and polyamides (various nylons). , acetalized polyvinyl alcohol (vinylon, etc.), polyacrylonitrile, polyethylene terephthalate, polyethylene, polypropylene, polyurethane, etc. Threads made of fine fibers or single fibers of synthetic organic polymers, natural fibers and regenerated cellulose, semi-synthetic or synthetic organic Examples include threads made of fibers mixed with polymer fibers. Various types of yarn can be used, including filament yarn, spun yarn, twisted yarn obtained by twisting these yarns, and composite yarn of filament yarn and short fibers.

Among these, twisted filament yarns are preferred. The thickness of the thread of the woven fabric is approximately 203 expressed in cotton spun yarn count.
to about 15O3, preferably from about 4O3 to about 12O3, or from about 35D to about 300D, preferably from about 45D to about 130D, in silk thread denier, the thickness of the woven fabric ranges from about 1100I1 to about 1100I1. Approximately 50
0μm preferably in the range of about 120n to about 350pm,
The porosity of the woven fabric ranges from about 40% to about 90%, preferably from about 50% to about 85%.

The woven fabric used for the porous development layer is a woven fabric that has been subjected to a degreasing treatment such as washing with water to substantially remove at least the oils and fats supplied or attached during the manufacturing of the yarn or textile.
Further, the woven fabric is subjected to a physical activation treatment (preferably glow discharge treatment, corona discharge treatment, etc.) disclosed in JP-A No. 57-66359 on at least one side of the fabric, or Showa 57-663
Hydrophilic treatment such as impregnation treatment with woody polymer disclosed in No. 59 et al., or sequential implementation of these treatment steps can make the textile hydrophilic and improve the adhesive strength with the lower layer (the side closer to the support). Can be strengthened.

For adhesion and lamination of a woven fabric on a reagent layer, water absorption layer or adhesive layer, JP-A-55-164356 and JP-A-57-6635 are used.
9 etc. can be followed. That is, water (or water containing a small amount of surfactant) is supplied substantially uniformly to the reagent layer, water absorption layer, or adhesive layer while it is still wet after application or after drying to swell the layer, and then the fabric is coated. The fabric is adhered and laminated onto the wet or swollen layer while applying light pressure evenly with a hand to integrate the fabric. When the hydrophilic polymer binder of the reagent layer, water absorption layer or adhesive layer is gelatin or gelatin derivative, a method is used in which the textile fabric is adhesively laminated and integrated while the gelatin (derivative) is in an undried gel state after application of the layer. Can be adopted.

In this way, an integrated multilayer analytical element with a textile spread layer is completed.

The fabric development layer of the integrated multilayer analytical element of the present invention is
Similar to the integral multilayer analytical element disclosed in US Pat.

In particular, when a reagent composition includes a dissociating agent for a test component bound to a polymer substance or protein, such as an enzyme, it may be preferable to include the reagent component in the textile spreading layer. . In order to contain the reagent composition or some components thereof in the textile spreading layer, rRese for the enzyme is used.
arch Disclosure J#12626 (Oc.
tober 1974), Japanese Patent Publication No. 50-137192,
It can be carried out according to the methods disclosed in JP-A-57-208997, etc. for enzyme substrates, JP-A-57-171,864, etc. for dissociating agents for test components bound to proteins, etc. can.

In addition to the above-mentioned layers, the integrated multilayer analysis element of the present invention may include JP-A-51-40191 and JP-A-53-13 as necessary.
Detection layer or mordant layer disclosed in JP-A-1089 etc., JP-A-1089-
Migration prevention layer disclosed in No. 29700 etc., JP-A No. 5
1-40191 etc., JP-A-56-854
Reagent-containing fine particle dispersion layer disclosed in No. 9, JP-A-52-348
A barrier layer consisting of a homogeneous thin layer of a specific hydrophobic polymer disclosed in JP-A-58-77661. Unexamined Patent Publication 1986-44
Air barrier layer disclosed in No. 865 etc., JP-A No. 60-214
A gas permeable adhesive intermediate layer disclosed in No. 52 can be incorporated therein.

The spreading layer, reagent layer, water absorption layer, detection layer, adhesive layer, etc. may contain a surfactant, preferably a nonionic surfactant. Examples of nonionic surfactants include p-octylphenoxypolyethoxyethanol, p-nonylphenoxypolyglycidol, polyoxyethylene oleyl ether, polyoxyethylene sorbitan monolaurate, oxytyl glucoside, and the like. By including a nonionic surfactant in the reagent layer, water absorption layer, or detection layer, water in an aqueous liquid sample is easily absorbed substantially uniformly into the reagent layer, water absorption layer, or detection layer during analysis operations, and Liquid contact with the spreading layer is rapid and substantially uniform. By containing a nonionic surfactant in the spreading layer, the spreading action (metering action) of the aqueous liquid sample becomes better. The content of the nonionic surfactant in the spreading layer is in the range of about 10 g to about 3.0 g, preferably about 20 g to about 2.0 g per rrf of the spreading layer.

The multilayer analytical element of the present invention can be prepared by known methods as described in the aforementioned patent specifications.

The multilayer analysis element of the present invention is cut into small pieces such as squares of about 15 to 30 squares on a side or circles of approximately the same size. It is preferable to use it as a chemical analysis slide by placing it in a slide frame as described in Japanese Patent Publication No. 58-32350, Japanese Patent Application Publication No. 58-501144, etc. from various viewpoints such as manufacturing, packaging, transportation, storage, and measurement operations. Depending on the purpose of use, it can be used in the form of a long tape and stored in a cassette or magazine, or small pieces can be attached or stored in a card with an opening.

The multilayer analytical element of the present invention can analyze analytes in liquid samples by the operations described in the aforementioned patent specifications. For example, an aqueous liquid sample such as whole blood, plasma, serum, urine, etc. in the range of about 5 pg to about 30 al, preferably 8 μe to 15 pffi, is spotted on the developing layer, and the sample is heated at about 20°C for 1 to 10 minutes. Incubation at a substantially constant temperature in the range of about 40°C, preferably around 37°C, causes color development or discoloration within the element to occur at wavelengths of maximum absorption of visible or ultraviolet light or The content of the analyte in the liquid sample can be determined by performing reflection photometry from the side of the light-transmitting support using light with a wavelength in the vicinity thereof, and using the principle of colorimetric measurement using a calibration curve prepared in advance. Alternatively, the analyte content in the liquid sample can be determined by photometrically measuring the intensity of fluorescence within the element and using a calibration curve prepared in advance. By keeping the amount of liquid sample spotted, incubation time, and temperature constant, quantitative analysis of analytes can be performed with high precision. Measurement operation is based on Japanese Patent Application Laid-Open No. 60-125543.
, JP-A-60-220862, JP-A-61-29436
7. Highly accurate quantitative analysis can be carried out with extremely easy operation using the chemical analyzer described in JP-A-58-161867 and the like.

The integrated multilayer analysis element of the present invention has a structure in which a woven fabric is provided as the uppermost layer as a porous layer (patch) for spotting and supplying an aqueous liquid sample with a certain shape and a certain area as disclosed in Japanese Utility Model Application No. 57-42951. You can also.

[Effect]

By using a fabric in which at least one of the weft and warp yarns is composed of a plurality of threads in the spreading layer, a spotted sample, even a whole blood sample, can be spread uniformly into a circular shape.

Hereinafter, the present invention will be specifically explained in detail with reference to Examples and Comparative Examples.

[Example] Example 1 Glucose quantification was carried out so that the coating amount of each component in the layer was as shown below on a colorless transparent polyethylene terephthalate (PET) film with a thickness of 180 μm that had been subjected to an undercoat treatment for gelatin. A reagent layer was applied.

Glucose oxidase 10000 U/I
Tf peroxidase 10000 U
/Bo 1,7-hydroxynaphthalene 100OQg
/rr(4-aminoantipyrine 500
tag/bolkali treated gelatin 20g
/rrf Nonylphenoxypolyethoxyethanol (contains average 10 oxyethylene units) 500 mg/r
rrA light shielding layer was further coated on this reagent layer so that the coating amount was as shown below.

Titanium oxide fine powder 10 g/rI?
Alkali-treated gelatin 5 g/n (nonylphenoxypolyethoxyethanol (contains an average of 10 oxyethylene units) 200 mg/rd; further, an adhesive layer was applied thereon to have the following coating amount to prepare a sheet for glucose determination. .

Alkali-treated gelatin 2.0 g/rd
Nonylphenoxypolythoxytoxyethanol (contains 10 oxyethylene units) 100 mg/n (separately, as wefts, 300 times/m of polyester filament 75d/36f whose main component is polyethylene terephthalate)
Two yarns twisted at 300 turns/m were used as the warp, and the warp was a polyester filament 75d/36f, which also has polyethylene terephthalate as its main component, twisted at a rate of 300 turns/m, with a weft density of 85 Z inches. A plain woven fabric having a warp density of 200 threads/inch was prepared. The fabric was treated with NaOll aqueous solution at about 95°C to
After % reduction processing, it was made hydrophilic by low-temperature plasma treatment and used as a fabric for a liquid sample spreading layer.

The previously prepared sheet for glucose determination was wetted almost uniformly with an aqueous solution of 0.2% nonylphenoxypolyethoxyethanol (containing an average of 10 oxyethylene units) and immediately brought into close contact with the fabric for the liquid sample spreading layer. Both were uniformly laminated by passing between pressure rollers. The obtained laminate was firmly adhered without peeling off even after drying.

In this way, an integrated multilayer analytical element for glucose determination was obtained.

Example 2 A glucose determination plate was prepared in the same manner as in Example 1, except that polyester filament 75d796f containing polyethylene terephthalate as the main component was used as the porous spreading layer, and a plain woven fabric was used in the same manner as in Example 1. We obtained body shape multilayer analysis elements.

Comparative Example 1 As a porous spreading layer, a single yarn of polyester filament 75d/36f whose main component is polyethylene terephthalate for both the weft and warp was twisted 300 times and 7m, and the weft density was 170 finches and the warp density was 200
An integrated multilayer analytical element for glucose determination was obtained in the same manner as in Example 1 except that a plain weave woven at a rate of 0/inch was used.

The following two types of measurements were carried out on the body type multilayer analysis element for glucose determination obtained as described above.

■Comparison of blood spreadability Whole blood collected using EDTA-2K (ethylefudiaminetetraacetic acid dipotassium salt) as a blood coagulation inhibitor was
Centrifuge at 250 rpm for 10 minutes under cooling at °C, mix blood cells and plasma so that the hematocrit value (Hc t) after reconstitution becomes the value shown in Table 1. Blood spreadability of integrated multilayer analytical element for glucose determination. Used for comparative testing.

■ Comparison of glucose quantitativeness Normal human whole blood containing approximately 10 On+g/dL glucose (B
et44%), the analytical operation for determining the glucose measurement value using three types of integrated multilayer analytical elements for glucose determination was repeated 20 times for each element.

In the above performance evaluation test, 10 pIl of sample (whole blood and reconstituted whole blood) was spotted on the spread layer of the element,
After incubation at 37°C for 6 minutes, the color developed in the element was immediately converted into an optical density value using visible light with a center wavelength of 540 na+ from the transparent support side.

The measurement results for the above two items are as shown in Tables 1 and 2.The integrated multilayer analytical element of the present invention using a woven fabric as a spreading layer in which at least one of the warp and weft is made of multiple threads has improved blood spreadability and It can be seen that the glucose concentration quantification is excellent.

■Blood spreadability evaluation results (Table 1) The body type multilayer analysis element for glucose determination shown in Examples 1 and 2 has a hematocrit value (Hc t
) changes, the shape of the spread is almost constant and close to a circle, but in the comparative example, when the Hct value of the blood to be spread changes, not only the spread area but also the shape of the spread changes. It can be seen that the integrated multilayer analytical element for glucose determination has better blood spreadability.

Table 1 Comparison of Blood Spreadability ■ Glucose Concentration Measurement Quantitative Evaluation Results The results in Table 2 are the measurement results obtained using the integrated multilayer analytical element for glucose analysis of Example 1.2 and Comparative Example 1.

Glucose concentration has been measured as being close to that of
Regarding measurement variation (simultaneous reproducibility), the integrated multilayer analytical element for glucose determination shown in Example 1 or 2 has better reproducibility than the one shown in Comparative Example 1, and provides highly reliable measurement results. I know that I can get it.

Table 2 Comparison of Glucose Quantitativeness * Coefficient of Variation [Effects of the Invention] The integrated multilayer analysis element of the present invention can uniformly spread a spotted liquid sample, and eliminates variations due to over-development in one direction. The reproducibility of the analysis results is good. In particular, even whole blood samples can be developed uniformly.

Patent applicant: Fuji Photo Film Co., Ltd. Unitika Co., Ltd.

Claims (2)

[Claims] (1) In an integrated multilayer analysis element formed by laminating in this order at least one reagent layer containing at least one reagent in a binder and a liquid sample spreading layer made of a fabric, the fabric constituting the liquid sample spreading layer is An integrated multilayer analysis element characterized in that at least one of the weft and warp consists of a plurality of threads. (2) The integrated multilayer analytical element according to claim 1, wherein the reagent layer is provided on one surface of a light-transparent water-impermeable planar support.