JPS60115861A - Optical measuring method of liquid sample using analytical appliance of flat plate vessel form - Google Patents

Abstract

PURPOSE:To improve analyzing accuracy by utilizing the optical path parallel with the base of an analysis appliance to measure optically the liquid sample introduced into an absorption chamber after reaction in a reaction chamber. CONSTITUTION:An observation chamber 14 is provided with side wall parts 14a, 14b formed of transparent materials facing each other and fixed in relative position so as to supply a specified optical path. If the parts 14a, 14b are so constituted that the space therebetween indicates a specified optical path, the optical measurement to detect the absorption, scattering and reflection of light is particularly easily accomplished with high accuracy by utilizing the optical path (the optical path in the direction X-X) parallel with the base of the analysis appliance. The analyzing accuracy is thus improved.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an optical measurement method that utilizes an integrated flat container analytical instrument for quantifying analytes in liquid samples.

[Description of technical field and existing technology] Until now, there has been a method for quantitatively analyzing various components (hereinafter referred to as analytes) contained in liquid samples, especially biological fluids such as urine, saliva, and blood. Wet methods and dry methods are known. The wet method is a method that has been commonly used for a long time, and involves, for example, reacting an analyte with a separately prepared reagent in a liquid phase in a container such as a test tube to produce a detectable change such as a red color. is generated and this is called Jilt
This is done through the specified operations.

However, while the wet method has the advantage of being able to quantify with high precision, it has the disadvantage of requiring skill in the analytical operations and not being able to be carried out by anyone in an emergency. Seven attempts have already been made to implement the wet method using simple containers.

For example, in Japanese Patent Publication No. 46-25596.

An analytical test pack is disclosed which comprises a pouch-like container of a flexible polymeric material, a portion of which is provided with an analytical reagent storage compartment, and a test sample inlet tube.

This analytical instrument consists of a reaction chamber, a pouch-like container made of a flexible polymeric material, and a chamber (herein referred to as observation chamber) for detecting detectable changes such as coloration produced by the reaction. Therefore, since i-measurement is easily deformed, it is difficult to maintain a constant optical path when measuring transmitted light in this observation room, which requires a complicated measuring device for analysis operations. Therefore, this analytical instrument has a problem in that it is not possible to perform highly accurate foot analysis using a small and inexpensive measuring device with a simple configuration.

On the other hand, considering the problem described in , the reaction chamber and the observation chamber are in phase η
Integrated flat vessel analyzers for quantifying analytes in liquid samples have also been developed, which are arranged in such a way that liquid flow between them is possible.

Such a flat container-type analytical instrument is originally a fl.
Since it was developed with the aim of making it easier to use and analyze data, and is generally used as an analytical instrument, it is inevitably constructed as a small structure with a thickness of several millimeters. In an analysis operation using a conventional flat container type analytical instrument, the optical A111 determination of the liquid sample introduced into the observation chamber after the reaction in the reaction chamber is performed by taking the optical path in the thickness direction of the analytical instrument. According to the practice, the optical path used for optical measurement is too short in this case, so it is difficult to obtain a 7111I viscosity of IR/f″r.

On the other hand, in the dry method, a liquid sample 1 is introduced into a sheet-like analytical element (also called an analytical film) containing a reagent 1, and changes such as coloration occurring within the analytical element are detected. This is done by optically detecting and quantifying the analyte.

While the dry method has the advantage of easy analytical operation and the ability to automate the analysis with a small all-in-one device, it has the disadvantage that it is difficult to achieve sufficient analytical accuracy t'+ depending on the analytical system. In particular, blood ZnS Oa# turbidity test
(ZTT) and blood oil. Timor J test (TTT)
), the dry method is not suitable for the purpose of quantifying the turbidity formed by the analyte in a liquid sample4. Since this method is carried out by taking an optical path, there is also a problem in that the optical path used for optical measurement is too short, making it difficult to obtain sufficient measurement accuracy.

[Summary of the Invention] The present invention is particularly directed to a measurement method using an integrated flat container-type analysis instrument for heavy machinery to analyze analytes such as biological fluids. The purpose of this invention is to provide an optical measurement method that contributes to the improvement of

The present invention uses an integrated plate-shaped container-type analytical instrument for limiting the analyte in a liquid sample, in which a reaction chamber and an observation chamber are connected so that liquid can flow between them. In the analysis operation, optical measurement of a liquid sample introduced into an observation chamber after a reaction in one reaction chamber is carried out using an optical path parallel (including substantially parallel) to the bottom surface of the analytical instrument. It consists of a method.

In other words, in an optical measurement method in an analytical method using a flat container-type analytical device, the optical path is taken in a direction parallel to the bottom surface of the analytical device rather than in a direction perpendicular to the plane of the analytical device, resulting in a longer optical path. can be set, the optical measurement according to the present invention improves the analytical accuracy.

[Detailed Description of the Invention] As a flat container type analysis instrument that can be advantageously used in carrying out the present invention, an analysis instrument having at least one of the following characteristics can be mentioned.

(I) By constructing at least a portion of the wall of the reaction chamber from a flexible material and housing a porous material exhibiting elasticity within the reaction chamber, stress is intermittently applied from the outside to the flexible wall portion of the reaction chamber. When applied, the flexible wall portion and the porous body are bent so as to exhibit resilient deformation in cooperation with each other.

(II) The sides of the observation room are provided with opposing wall sections made of transparent material whose relative positions are fixed and which provide a constant light path.

(III) There is no substantial partition between the reaction chamber and the observation chamber regarding the flow of liquid.

An example of a plate-shaped container-type analysis device having a configuration including all of the above-mentioned features is shown in the accompanying drawings.

The present invention will be described in detail below, taking as an example an analysis method using a flat container-type analysis instrument illustrated in the accompanying drawings.

Figure 1-A is a perspective view showing an example of the configuration of a flat container-type analytical instrument, and Figure 1-B is a perspective view taken along line 1-1 of the analytical instrument in Figure 1-A. FIG. However, the first
In Figure A, the analytical instrument is shown with the upper sheet-like wall (lid) portion removed.

Analyzer Jl shown in Figures 1-A and 1-B
l Q is polyethylene, polypropylene, polystyrene, polymethacrylate ester resin.

Hard materials such as polyacrylate resin, bisphenol A polycarbonate, polyvinyl chloride, etc.
Alternatively, it is composed of a flat container part 11 made of a synthetic resin material that is relatively difficult to deform, or a glass plate, and an upper sheet-like upper wall part 12, and is in the form of a flat container as a whole. Inside the container section 11, a reaction chamber 13 and an observation chamber 14 are connected to each other so that liquid can flow therebetween. A porous body 15 having elasticity is housed inside the reaction chamber 13. As the porous body used for this purpose, for example, a structure made of a porous resin material such as polyamide, HIi acetate fibrous resin, polyvinyl chloride, polyethylene, polypropylene, polyurethane, etc. can be used. Alternatively, woven lIi, non-woven/I+, felt, paper, or a composite material of these materials and a porous resin structure can also be used.

In addition, considering the deformation function described later, the porous body has a sea I
・It is desirable that the shape be the same.

In addition, in FIG. 1-A, the sea I-shaped wall portion 12 of the -L portion is shown in a separated state, but this sea-shaped wall portion 12 is actually shown in FIG. 1-B. As shown, it is attached to the container part 11 in a watertight manner to form an integrated flat container-type analysis instrument 10.

The sheet-like upper wall portion 12 is made of synthetic resin such as polyethylene terephthalate or polyamide, various elastomers, metal such as aluminum, copper, stainless steel, or a similar flexible material, and is a flexible material that exhibits flexibility when formed into a thin sheet. It is in the form of a relatively thin sheet made of a material (particularly preferably an elastomer exhibiting self-restoring properties or a composite material of an elastomer and a metal sheet), and the sheet-like −L portion wall portion 12 is not subjected to external stress. It is designed so that it can be easily deformed when it is damaged. Therefore, when stress is applied intermittently from the outside to the sheet-like upper wall portion 12 of the reaction chamber, the flexible sheet-like two-part wall portion 12 and the porous body 15 cooperate to improve the resilience. The deformation that exists will be repeatedly reduced. This resilient deformation caused by the cooperation between the sheet-like upper wall 12 and the porous body 15 enables sufficient mixing of the liquid reagent introduced into the reaction chamber and the reagent by repeating the deformation. I+(
make it possible. In addition, in the explanation so far, the sheet-like sheet 1 i
Although the example in which the ′:r′ portion 12 is made of a flexible material has been shown, the sheet-like two-part wall portion 12 may be made of a hard material that hardly exhibits physical deformation, or may be made of a thick wall. On the other hand, the bottom of the container portion 11 may be made of a flexible material. Alternatively, both the sheet-like upper wall portion 12 and the bottom of the container portion 11 may be made of flexible material.

In addition, when a material with high self-restoring properties is used as the above-mentioned flexible material, such as elastomer sheet 1 or a composite material of an elastomer sheet and a thin metal sheet, the porous material described in 1-1. It is also possible to omit its use.

A liquid sample is generally introduced into the reaction chamber 13 through a sample inlet 16 provided in the sheet-like wall 12 . As the liquid sample, blood (whole surface, plasma, or serum) can be used as it is, or a diluted liquid sample can be used. There are two methods for diluting the liquid sample: one is to dilute the liquid sample with a diluent in advance, and the other is to separately introduce the liquid sample 1 and the diluent (the latter can be determined as appropriate) into the reaction chamber. It is possible to select and implement the method as appropriate. A reagent that directly or indirectly causes a detectable change such as coloration by reacting with the analyte contained in the liquid sample must be stored in the reaction chamber 13 in advance, or the reagent must be stored in the reaction chamber 13 in advance, or , Jl 6 into the reaction chamber 13 from the analytical operation surface. However, in order to quickly and reliably react the analyte and reagent in the reaction chamber 13, it is necessary to impregnate the reagent into the porous body in advance, apply the reagent to the inner wall of the reaction chamber, or attach it by printing, etc. It is preferable to store it in The reagent used may be in a dry state (solid), semi-dry state (jelly-like, etc.), or liquid. Solid sample is T
(It may be in the form of i-grains or powder. The sample injection port 16 is normally closed by the sample injection port lid 18 after the sample is introduced into the reaction chamber 13.

The observation chamber 14 receives a sample solution exhibiting a detectable change such as coloration caused by contact between the analyte and the reagent in the reaction chamber 13, and detects the change by optical means. This is the part used to perform Analyte's Release 1 j, j.

The observation chamber 14 is preferably provided with opposing side wall portions 14a, 14b of transparent material whose relative positions are fixed and which provide a constant light path. If the space between these transparent side wall portions 14a and 14b is configured to indicate a constant optical path, the optical path will be directed in the conventional longitudinal direction, that is, in a direction perpendicular to the plane of the flat analytical instrument IO. Instead of an optical measurement method with an optical path set parallel to the bottom of the analyzer (e.g.
By using the x-direction optical path), optical measurements for detecting absorption, scattering, reflection, etc. of light can be performed particularly easily and with high precision. The optical path does not need to be strictly in line 11a at the bottom of the observation room, nor does it need to be perpendicular to the side wall.

Regarding the flow of liquid between the reaction chamber 13 and the observation chamber 14,
It is desirable that there be no substantial partition walls. This means, for example, that between the reaction chamber 13 and the observation chamber 14, normal artificial pressure from the outside (for example, pressure applied to the reaction chamber by the hands of an analytical operator or by means of a Jllll constant apparatus) is applied. septum function (such as suppression) that cannot be controlled by
For example, capillary action, dialysis action through semipermeable membranes, etc.) are not provided, and this requirement makes it difficult to observe the mixing operation of the liquid sample and reagent introduced into the reaction chamber. Since the chamber also functions as part of the reaction chamber,
There is an advantage that the desired mixing can be carried out easily and sufficiently. Another advantage is that the sample solution can be quickly moved to the observation chamber after the necessary analyte and reagent have come into contact with each other, making it easier to speed up the analysis operation.

However, it is also possible to install a net or a filter sheet between the reaction chamber and observation chamber to prevent the movement of solids, as long as it does not impede substantially free liquid flow between them. It is.

In the flat container-type analytical instruments shown in Figures 1-A and 1-8, an example of a configuration in which the reaction chamber and observation chamber are substantially the same and located on the 1L plane is shown. , the positional relationship between the reaction chamber and observation chamber is not limited to this configuration; for example,
As shown in FIG. 2, the reaction chamber and the observation chamber can also be arranged in a stacked manner.

Figure 2 shows a reaction chamber 24 and an observation chamber 25 arranged in a stacked manner.
It is a figure which shows the longitudinal cross-sectional view of the flat container type analytical instrument 20 of the form which consists of.

As in the case of FIGS. 1-A and 1-8, the analytical instrument 20 is composed of a flat container portion 21 and a sheet-like upper wall portion 22 made of a partially flexible material. ,
It is in the form of a qi plate-shaped container as a whole. Container part? Inside (2), a reaction chamber 23 and an observation chamber 24 are connected in a stacked manner through an opening 27 so as to allow substantially free fluid flow between them. A porous body 25 exhibiting elasticity is housed inside the reaction chamber 23 .

The analysis operation using the analysis instrument in the form of a laminate shown in FIG. 2 is almost the same as in the previous example. That is, after introducing a liquid sample from the sample injection port 26, the sample injection port 26 is closed with the sample injection port lid 28, and in the reaction chamber 23, intermittent pressure is applied from the outside to the flexible sheet-like upper wall portion 22. By repeatedly bringing about deformation exhibiting restorability in cooperation with the elastic porous body 25, the liquid sample and reagent are sufficiently mixed to cause a reaction between the analyte and the reagent, and then the sample liquid is transferred to the observation chamber 24. A method is used in which optical measurements are performed by moving the

In the analysis instrument 2o shown in FIG. 2, the optical path is directed in a direction parallel or approximately parallel to the bottom of the analysis instrument 2o (for example, in the This makes it easy to set the analyzer in the perpendicular direction, and it is possible to secure a long optical path for optical measurements, thereby easily achieving an improvement of one analyzer degree.

In the explanation so far, the four features and configuration of the present invention have been shown by taking the plate-shaped container type analytical instrument shown in the attached drawings as an example, but when implementing the present invention, it is necessary to use a configuration different from such a configuration. Of course, it is possible to use a variety of plate-shaped container-type analysis instruments having various configurations.

There are no particular limitations on the samples and analytes contained in the samples that can be analyzed using the flat container type analytical instrument, and various types of samples that are subject to conventional analytical operations such as wet or dry methods. samples and analytes can be analyzed.

Examples of such analysis systems include the following analysis systems.

i) A porous body was made to contain barbiturate and barbiturate sodium, and after being washed with an aqueous zinc sulfate solution, it was placed in a reaction chamber, and then serum (liquid sample) was introduced into the reaction chamber and thoroughly stirred. after.

A system that calculates the ZTT value in a liquid sample by optically measuring the sample solution at a measurement wavelength of 660 nm in an observation room.

11) An aqueous solution containing thymol, barbiturate, and parbital sodium is applied to the wall of the reaction chamber, allowed to dry, and placed in the reaction chamber. Serum (liquid sample) is introduced into the reaction chamber and mixed thoroughly, and then the sample solution is added. In the observation room, the transmitted optical density at the optical path length is determined using visible light with a center wavelength of 660 nm, and the TT and T values in the liquid sample are calculated using the colorimetric method.

111) Anti-B-lipoprotein antibody was contained in a porous material, this was housed in a reaction chamber, and a liquid sample containing B-lipoprotein was introduced into the reaction chamber and thoroughly stirred to allow the immune reaction to proceed. This system then quantifies the B-lipoprotein content in a liquid sample by optically measuring the resulting turbid liquid in an observation room.

1t) An aqueous solution containing ADP, AMP, creatine phosphate, glucose-6-phosphate, dehydrogenase, glucose, hexokinase, and NAD is applied to the wall surface of the reaction chamber, dried, and stored in the reaction chamber. Introduce ion-distilled water into the reaction chamber and allow it to warm up sufficiently, then measure the transmitted optical density at the optical path length of the solution containing NADH produced by creatinine kinase in serum in the observation chamber using near-ultraviolet light with a center wavelength of 340 nm. The NADH is
Or a system for quantifying creatine phosphokinase.

[Brief explanation of drawings]

FIG. 1-A is a perspective view showing an example of the configuration of a flat container-type analytical instrument that can be used for carrying out the present invention, and FIG.
Figure 1-B is a longitudinal sectional view taken along line I-I of the analytical instrument of Figure 1-A. However, in FIG. 1-A, the analytical instrument is shown with its two sheet-like wall (lid) parts removed. FIG. 2 is a diagram showing an example of another configuration of a flat container-type analytical instrument that can be used to implement the present invention. 10.20.30: Flat container type analytical instrument 11.21.
31: Container part 12.22.32: Sheet-like 1tt part 13 of J2 part
．． 23.33: Reaction chamber 14.24.34: Observation chamber 15.25.35: Porous body 16.26.36: Sample injection 18.28: Sample injection Self-attachment 27: Opening Patent applicant Fuji Photo Film Co., Ltd. Agent Patent Attorney Yasuo Yanagawa

Claims (1)

[Scope of Claims] 1. An integrated flat plate for determining an analyte in a liquid sample, the reaction chamber and the observation chamber being connected and arranged such that liquid communication between them is possible. In the operation of performing analysis using a container-type analysis instrument, the optical path of the liquid sample introduced into the observation chamber after the reaction in the reaction chamber (fy measurement) is characterized by using an optical path parallel to the bottom of the measurement analysis instrument. 2. Using a flat container-type analysis instrument in which at least two opposing side walls of the observation chamber are made of a hard transparent material, thereby forming a fixed optical path. The optical measurement method according to claim 1, characterized in that a plate-shaped container-type analytical instrument is used in which the reaction chamber and the observation chamber are substantially on the same plane. The optical measuring method according to claim 1. The optical measurement method according to claim 1, characterized in that a flat container-type analysis instrument having a 4° reaction chamber and an observation chamber arranged in a stacked manner is used. An optical measurement method of 5゜ flat container-type analytical instrument, in which at least a part of the wall of the reaction chamber is made of a flexible material that exhibits self-righting properties, and the side of the observation chamber is
Opposed wall sections of transparent material whose relative positions are fixed to provide a constant light path are provided, and between the reaction chamber and the observation chamber there is a substantial separation with respect to liquid flow. The optical measuring method according to claim 1, characterized in that there is no partition wall. The 6゜flat container-type analytical instrument has at least a portion of the wall of the reaction chamber made of a flexible material, and a porous material exhibiting ionic properties is accommodated in the reaction chamber, so that the flexible wall of the reaction chamber is When stress is intermittently applied to the portion from the outside, the flexible wall portion and the porous body exhibit resilient deformation in association with i+li. The sides of the observation chamber are equipped with opposing wall sections made of transparent material whose relative positions are fixed and which provide a constant light path, and between the reaction chamber and the observation chamber there are Claim 1-1 Optical measurement method 2, characterized in that there is no substantial partition wall regarding the flow of liquid.