JPH08226921A - Reagent container and measurement method using the container - Google Patents

Abstract

PURPOSE: To always completely shield reagent from an atmosphere for allowing the reagent to be preserved and used for a long time by providing a flexible container or a bulkhead which moves or deforms in the container and reducing an internal volume of the container according to a decrease in the amount of housed reagent. CONSTITUTION: Reagent is housed into a container formed of flexible material such as synthetic resin or a metal foil or into a container having a movable or flexible bulkhead in it. When the reagent in the container is sucked out of the container by a tube attached in a sealed state to the container, the inside of the container is at a negative pressure, so that the flexible container deforms while in the case of the container having the bulkhead the bulkhead deforms or moves to have its internal volume reduced. Therefore, since an atmosphere is prevented from flowing into the container as the reagent is used and the reagent is always completely shielded from the atmosphere, the reagent can be stably preserved without deterioration in performance, thereby permitting correct measurement.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a reagent storage container capable of stably storing a stored reagent for a long period of time and a method for measuring a test substance using the reagent storage container. In the present invention, by blocking the stored reagent from the outside air, deterioration of the performance of the reagent can be prevented, and the reagent can be stored and used for a long period of time. INDUSTRIAL APPLICABILITY The present invention is particularly suitable for long-term storage and use of a reagent such as a clinical test drug, an analytical reagent, or a standard solution that is easily deteriorated and difficult to store.

[0002]

2. Description of the Related Art Conventionally, reagents such as clinical test agents, analytical reagents, and standard solutions are glass, polyethylene, polyvinyl chloride for handling or storing at each stage of production, distribution, sale, use or storage. A container made of synthetic resin such as, for example, which cannot be deformed and whose internal volume cannot be reduced in accordance with a decrease in its internal volume, or a container which can be deformed but whose internal volume cannot be reduced in accordance with a decrease in its internal volume. Has been stored in and handled.

By the way, these conventional containers cannot be deformed and the internal volume cannot be reduced in accordance with the decrease of the internal volume, or even if the container is deformable, the internal volume is reduced in accordance with the decrease of the internal volume. In the case of a container that cannot be used, when the necessary amount of the reagent stored in the container is transferred to the outside of the container and used, an amount of outside air corresponding to the amount of the reagent transferred to the outside of the container flows into the container. The carbon dioxide contained in the air, which is the outside air, dissolves in the reagent remaining in the container, and the pH of the reagent
And the performance as a reagent is deteriorated.

For example, a reagent for measuring alkaline phosphatase (ALP) which is a clinical test agent, a reagent for measuring leucine aminopeptidase (LAP), a zinc sulfate turbidity test (ZT)
T, Kunkel reaction) measuring reagent, thymol turbidity test (TT
For T) measuring reagent, magnesium (Mg) measuring reagent, calcium (Ca) measuring reagent, etc., because carbon dioxide in the outside air dissolves in the reagent during storage of the reagent and the pH of the reagent decreases, The measurement reaction was disturbed and an error occurred in the measured value, and it was uneconomical to store and use for a long period of time.

Further, the oxygen contained in the air, which is the outside air flowing into the container, dissolves in the reagent remaining in the container to oxidize the constituent components of the reagent, deteriorating the performance as a reagent.

Further, the reagent used in the analyzer is placed in the analyzer or a position adjacent thereto, and the reagent is sucked and used through a tube inserted in the container, or is aspirated by a pipette for dispensing. It will be poured and used. Therefore, the container in which the reagent is stored cannot be hermetically sealed in order to pass the tube or to put in and take out the pipette for dispensing, and the outside air is always flowing into the container. Therefore, it is more likely to be affected by the above carbon dioxide or oxygen, and the deterioration of the performance as a reagent is promoted.

In addition to this, the evaporation of water causes the constituent components in the reagent to be concentrated, which causes an error in the measured value. In particular, the error caused by this concentration was fatal in the standard solution, the standard serum and the like, which serve as a standard for calculating an accurate measured value. As described above, the containers conventionally used for storing the reagents and the method for measuring the test substance using these containers have various problems.

[0008]

As described above, the conventional container cannot be deformed and the internal volume cannot be reduced according to the decrease of the internal volume, or even if the container is deformable, the internal volume can be reduced according to the decrease of the internal volume. In a container whose internal volume cannot be reduced,
An amount of outside air corresponding to the amount of reagent used flows into the container, carbon dioxide contained in the outside air dissolves in the reagent, and the pH of the reagent is lowered, or the amount of outside air contained The oxygen contained in the reagent oxidizes the constituent components of the reagent, which causes errors in the measured values and makes it difficult to store and use for a long time, which is uneconomical.

Further, the container for storing the reagent, which is placed in the analyzer or in the vicinity thereof, cannot be hermetically sealed in order to pass the tube or put in and take out the pipette for dispensing,
Outside air is always flowing into the container. Therefore, it becomes more susceptible to the above-mentioned carbon dioxide or oxygen,
The deterioration of the performance as a reagent is promoted, and further, the components in the reagent are concentrated due to the evaporation of water,
Therefore, an error occurs in the measured value. In addition, since it is not hermetically sealed in this way, microorganisms such as bacteria and mold may mix and propagate in the container, degrading the performance of the reagent.

The inventors of the present invention have made extensive studies aiming at eliminating the disadvantages of the conventional containers used for storing reagents and the methods for measuring a test substance using these containers. By storing the reagent in the reagent storage container whose internal volume decreases in accordance with the decrease in the amount, it is possible to always completely shield the reagent from the outside air through each stage of distribution, sale, use and storage of the reagent, And it was found that it does not hinder its use, it is not affected by carbon dioxide or oxygen in the air, which is the outside air, and it is possible to prevent the concentration of constituent components in the reagent due to evaporation of water, and to prevent contamination by microorganisms. Completed a reagent storage container that can prevent breeding, can store reagents for a long time and can be used, and a method for measuring a test substance that can perform accurate measurements for a long time. It was.

[0011]

The present invention includes the following inventions. (1) A reagent storage container in which the internal volume decreases as the internal volume decreases. (2) The reagent storage container according to (1) above, wherein the reagent storage container is a deformable container that is molded so as to be hermetically sealed. (3) The reagent storage container according to (1), wherein the reagent storage container has a partition wall that is movable or deformable in the container and is molded so as to be hermetically sealed. (4) A method for measuring a test substance using a reagent storage container whose internal volume decreases as the internal volume decreases. (5) The method for measuring a test substance according to (4), wherein the reagent storage container is a deformable container that is molded so as to be hermetically sealed. (6) The method for measuring a test substance according to (4), wherein the reagent storage container has a partition wall that is movable or deformable in the container and is formed so as to be hermetically sealed.

The reagent storage container according to the present invention has an internal volume that decreases in accordance with a decrease in the internal volume. Therefore, when the required amount of the reagent stored in the container is transferred to the outside of the container and used, By reducing the internal volume corresponding to the amount of the reagent transferred to, the outside air can be prevented from flowing into the container, and the reagent in the container and the outside air can be completely shut off.

In the reagent storage container of the present invention, the decrease in the internal volume in accordance with the decrease in the internal volume can be achieved, for example, by using a deformable container. In a deformable container, when the reagent stored in the container is sucked and transferred to the outside of the container, the inside of the container becomes a negative pressure and the container is deformed accordingly, which causes the internal volume to decrease according to the decrease in the internal volume. Can be reduced.

It should be noted that in a container that cannot be deformed and whose internal volume cannot be reduced in accordance with a decrease in its internal volume, or if it is deformable but whose internal volume cannot be reduced in accordance with a decrease in its internal volume, , When the reagent stored in the container is sucked and transferred to the outside of the container and the inside of the container becomes a negative pressure, the internal volume cannot be reduced in accordance with the decrease in the internal volume. When the container is sealed, a large negative pressure is generated inside the container, and it becomes difficult to suck and transfer the reagent stored in the container to the outside of the container.

It is also possible to achieve a decrease in the internal volume according to the decrease in the internal volume by using a container having a movable or deformable partition wall inside the container. In a container having a movable or deformable partition inside the container, when the reagent stored in the container is sucked and transferred to the outside of the container, the inside of the container becomes a negative pressure, and accordingly the partition moves or deforms. As a result, the internal volume can be reduced according to the decrease in the internal volume. Further, in the container having the movable or deformable partition wall in this container, the internal volume can be reduced in accordance with the decrease in the internal volume by actively moving or deforming the partition wall.

The shape and size of the reagent storage container in the present invention are not particularly limited, and any shape and size can be used.

In the reagent container of the present invention,
The material of the deformable container may be any material that can be deformed from the raw material of the material, such as polyethylene, polypropylene, polyvinyl chloride, fluororesin, epoxy resin, polyurethane resin, polyamide, ethylene vinyl acetate. One or more kinds of materials such as a copolymer, ethylene vinyl alcohol, polyethylene terephthalate, synthetic resin such as polyester or polyvinylidene, metal foil such as aluminum foil or carbon fiber can be used in combination.

The material of the container having a partition wall that is movable or deformable is polyethylene, polypropylene, polyvinyl chloride, fluororesin, epoxy resin, polyurethane resin, polyamide, ethylene vinyl acetate copolymer, ethylene. Vinyl alcohol, polyethylene terephthalate, polyester, polyvinylidene, polycarbonate, diallyl phthalate, urea resin, melamine resin, silicon resin, styrene resin, acrylic resin, polyacetal, vinyl acetate, synthetic resin such as phenol resin or polystyrene, metal, carbon fiber, One or more kinds of materials such as glass or ceramics can be used in combination.

The container for storing a reagent in the present invention can be obtained by manufacturing a container according to a known method such as heat molding using the raw material of the container such as the above-mentioned synthetic resin, metal, carbon fiber, glass or ceramics. it can.

For example, a bag-shaped container is formed by stacking deformable sheet materials made of the above-mentioned materials of a single layer or a laminated body and fusing or adhering the peripheral edges thereof by heat fusion or high frequency fusion. Can be manufactured. In this case, the reagent storage portion for storing the reagent is formed in the inner portion surrounded by the fusion-bonded portion or the adhesive portion on the peripheral edge.

In the deformable container, depending on the material of the container, if the thickness of the container wall is too thick, the deformation may be hindered. In such a case, the deformation is not hindered. It is necessary to select the thickness of the container wall.

The reagent storage container according to the present invention, when the necessary amount of the reagent stored in the container is transferred to the outside of the container for use,
By reducing the internal volume corresponding to the amount of reagent transferred to the outside of the container, the outside air is prevented from flowing into the container and the reagent inside the container and the outside air are completely cut off. It is necessary that outside air does not flow into the container even when it is not being transferred. In other words, it is necessary that the reagent in the container and the outside air can always be completely shut off. This may be formed so that the container can be sealed. Then, the opening of the container may be plugged or sealed.

When the reagent is stored in the reagent storage container of the present invention and placed in the analyzer or at a position adjacent thereto, and the reagent is sucked through a tube connected to the container and used for analysis In addition, the connection between the container and the tube must be sealed so that outside air does not flow into the container or the tube. This is to integrally mold the connection between the container and the tube, or to provide the container with a connection that allows the tube to be inserted closely,
The opening of the container and the tube can be connected to each other through a connection tool that can be closely connected to each other, or the connection between the container and the tube can be sealed with a sealing material or a sealing tape. it can.

Further, when the reagent is stored in the reagent storage container of the present invention and used by being placed in the analyzer or a position adjacent thereto, the reagent stored in the container is aspirated by the pipette for dispensing and dispensed. In case of use for analysis, the opening of the container must be able to pass the reagent suction part of the pipette for dispensing and must be sealed so that external air does not flow into the container. This can be achieved by, for example, sealing the opening of the container by using a material having elasticity at least the portion through which the reagent suction section of the pipette for dispensing passes.
It can be carried out. The portion of the reagent storage container of the present invention that faces the outside air preferably has a gas barrier property.

In the present invention, the term "reagent" means detection or quantification of a test substance by a chemical method, a biological method, a biochemical method, an immunological method, a molecular biological method, or the like, synthesis, preparation of the substance, It is a product used for purification or production or measurement of physicochemical properties of a substance. And
Examples of the substances used for detection or quantification of the test substance by this chemical method, biological method, biochemical method, immunological method, molecular biological method, etc. Examples include a measurement reagent for a test substance, a standard substance, a standard solution, a standard serum, a control serum, and the like.

In the method for measuring a test substance according to the present invention, which uses a reagent storage container whose internal volume decreases in accordance with the decrease in internal volume, the internal volume of the reagent decreases in accordance with the decrease in internal volume. The reagent is stored in a container for storing a reagent, and a test substance is measured using the stored reagent.

The test substance in the present invention is a chemical method,
It is a substance that is detected or quantified by a biological method, a biochemical method, an immunological method, a molecular biological method, or the like, or a substance whose physicochemical properties are measured.

Examples of substances containing the test substance include reagents, biological samples, meat, vegetables, grains, fruits, marine products, forest products, processed foods, beverages, drinking water, natural products or industrial products. it can.

Of these, examples of biological samples include human or animal blood, serum, plasma, urine, stool, semen, cerebrospinal fluid, saliva, sweat, tears, ascites, amniotic fluid, brain and other body fluids or organs, hair, Examples thereof include tissues or cells such as skin, nails, muscles and nerves.

The test substances include, for example, alkaline phosphatase (ALP), acid phosphatase (ACP), leucine aminopeptidase (LAP),
Cholinesterase (Ch-E), aspartate aminotransferase (AST, GOT), alanine aminotransferase (ALT, GPT), lactate dehydrogenase (LDH), γ-glutamyl transpeptidase (γ-GTP), creatine kinase (CK) , Α-
Amylase, total bilirubin, direct bilirubin, creatinine, creatine, urea nitrogen (BUN), uric acid (U
A), albumin, total protein, ammonia, total cholesterol, free cholesterol, HDL-cholesterol, triglyceride (TG), phospholipid (PL), free fatty acid (NEFA), glucose, sodium, potassium, chlor, inorganic phosphorus (IP) , Calcium (C
a), magnesium (Mg), serum iron (Fe), unsaturated iron binding capacity (UIBC), serum copper (Cu), zinc sulfate turbidity test (ZTT), thymol turbidity test (TTT), CR
P, immunoglobulin G (IgG), immunoglobulin A
(IgA), immunoglobulin M (IgM), rheumatoid factor (RF), anti-streptolysin O antibody (AS
O), complement C ₃ , complement C ₄ , lipoprotein (a) [Lp
(A)], α ₁ -acid glycoprotein, haptoglobin,
Fructosamine, HBs antigen, anti-HBs antibody, HBe antigen, anti-HBe antibody, anti-HBc antibody, anti-HCV antibody, anti-H
IV antibody, anti-ATLV antibody, viral DNA or RNA, bacterial DNA, hormones such as human placental chorionic gonadotropin (hCG), tumor markers such as CA19-9, blood coagulation system-related substances such as antithrombin III (ATIII) Fibrin decomposition products (FDP) and other fibrinolytic system-related substances, allergen-specific IgE antibodies, and the like.

In particular, alkaline phosphatase (AL
P), leucine aminopeptidase (LAP), zinc sulfate turbidity test (ZTT), thymol turbidity test (TTT),
When the carbon dioxide in the outside air, such as magnesium (Mg) and calcium (Ca), dissolves in the reagent and the pH of the reagent decreases, the measurement reaction is disturbed and the measured value has an error. It is suitable for the test substance that is generated.

As the measuring method of these test substances, for example, a chemical measuring method, a biological measuring method,
Examples thereof include biochemical measurement methods such as enzyme reaction, immunological measurement methods such as antigen-antibody reaction, and molecular biological measurement methods such as reaction between genes or fragments thereof.

[0033]

The container for storing a reagent according to the present invention has an internal volume which is reduced in accordance with a decrease in the internal volume, and when the required amount of the reagent contained in the container is transferred to the outside of the container and used, By reducing the internal volume corresponding to the amount of the reagent transferred to, the outside air can be prevented from flowing into the container, and the reagent in the container and the outside air can be completely shut off.

In the deformable container which is one of the reagent storing containers of the present invention, when the reagent stored in the container is sucked and transferred to the outside of the container, the inside of the container becomes negative pressure, and accordingly the container Is deformed, whereby the internal volume can be reduced in accordance with the decrease in the internal volume.

Further, in a container having a partition wall which is movable or deformable in the container which is another one of the container for storing the reagent of the present invention, when the reagent contained in the container is sucked and transferred to the outside of the container. The inside of the container has a negative pressure, and the partition moves or deforms in response to the negative pressure, whereby the internal volume can be reduced in accordance with the decrease in the internal volume. Further, in the container having the movable or deformable partition wall in this container, the internal volume can be reduced in accordance with the decrease in the internal volume by actively moving or deforming the partition wall.

[0036]

EXAMPLES The present invention will be described in more detail below with reference to examples, but the present invention is not limited to these examples.

Example 1 Storage stability test of reagent storage container whose internal volume decreases with decreasing internal volume and leucine aminopeptidase assay reagent stored in this container

A container for storing a reagent was prepared in which the internal volume was reduced in accordance with the decrease in the internal volume, and the storage stability of the leucine aminopeptidase (LAP) measuring reagent contained in this container was tested.

(1) Two 16 cm × 12 cm gas barrier sheets having a thickness of 0.1 mm and having low density polyethylene on both sides of a polyamide core were laminated. (2) Tetrafluoroethylene-hexafluoropropylene copolymer (FEP) having an inner diameter of 1 mm and a length of 60 cm
After inserting the tube made of 5 mm between the two sheets of (1), heat-sealing the periphery of this sheet with a width of 5 mm,
The bag-shaped reagent storage container A of the present invention (volume 200 m, whose internal volume decreases as the internal volume of the shape shown in FIG. 5 decreases)
L) was prepared.

[0040]

[Figure 5]

(3) Two 0.5 cm thick gas barrier sheets of 16 cm × 12 cm in which both sides of the core of ethylene vinyl alcohol are low density polyethylene were stacked. (4) Tetrafluoroethylene-hexafluoropropylene copolymer (FEP) having an inner diameter of 1 mm and a length of 60 cm
After inserting the tube made of 5 mm between the two sheets of (3), the peripheral edge of this sheet is heat-fused with a width of 5 mm,
The bag-shaped reagent storage container B of the present invention (capacity 200 m, whose internal volume decreases in accordance with the decrease in internal volume of the shape shown in FIG. 5)
L) was prepared.

(5) 50 mM Tris buffer (pH
8.0) was prepared and used as the first reagent of leucine aminopeptidase (LAP) measurement reagent. (6) Leucine aminopeptidase (LA) was prepared by preparing a reagent containing 20 mM Tris buffer (pH 6.0) containing 10 mM L-leucine-4-nitroanilide hydrochloride.
P) It was used as the second reagent of the measurement reagent.

(7) The first reagent of (5) and the second reagent of (6) are respectively placed in reagent storage containers A of (2) and (4).
Both of the reagent storage containers B of No. 1 were filled. (8) Reagent storage container A containing the first reagent of (7), reagent storage container A containing the second reagent, reagent storage container B containing the first reagent, and reagent containing the second reagent The storage container B was placed in a reagent storage of a Hitachi 736-15 type automatic analyzer maintained at a temperature of 10 ° C., and each of these tubes was connected to a reagent switching valve of this automatic analyzer. And saved.

(9) As a control, the outer diameter is 158 mm
(Depth, L) x 115 mm (height, H) x 22 mm
(Width, W) and rectangular polyethylene container having a 13 mm diameter opening on the upper surface (control container) (capacity 300
(5) the first reagent and (6) the second reagent, respectively. Next, a control container containing the first reagent and a control container containing the second reagent were stored in a reagent container in which the temperature of the Hitachi 736-15 type automatic analyzer was kept at 10 ° C as in (8). A tetrafluoroethylene-hexafluoropropylene copolymer (F having an inner diameter of 1 mm and a length of 60 cm, which was placed in a container and inserted into the control container through an opening (F
The other end of the tube made from EP) was connected to the reagent switching valve of this automatic analyzer. And saved.

(10) The stability of each of the reagents (8) and (9) stored in a reagent container at 10 ° C. was tested. Storage using the first reagent and the second reagent stored in the reagent storage container A, the first reagent stored in the reagent storage container B, the second reagent, and the first reagent and the second reagent stored in the control container, respectively. Start date, storage 2 days, 4 days, 6 days, 10
After 5 days, 12 days and 14 days, a serum sample having a leucine aminopeptidase activity value of 63 IU (international unit) / L was measured in quintuplicate. In addition, this Hitachi 7
The measurement with the 36-15 type automatic analyzer was carried out by adding 250 μL of the first reagent to 8 μL of the sample and incubating at 37 ° C. for 3 minutes, and then adding 125 μL of the second reagent and reacting at 37 ° C.
Second reagent addition at 15 nm and subwavelength 480 nm 1
It was performed by calculating the leucine aminopeptidase activity value from the rate of increase in absorbance from minute 18 seconds to 4 minutes 30 seconds.

The stability test results of this stored reagent are shown in FIG. In addition, in FIG. 1, the horizontal axis represents the number of days elapsed after the start of storage, and the vertical axis represents the relative activity when the leucine aminopeptidase activity values of five-fold measurement were averaged and divided by the leucine aminopeptidase activity value at the storage start date. Represents a value.

[0047]

[Figure 1]

From this figure, in the reagent stored in the reagent storing container A and the reagent storing container B of the present invention, 10
Even after 14 days of storage at 0 ° C, the same leucine aminopeptidase activity value as that of the storage start date could be exhibited, and accurate measurement could be performed, whereas it was stored in the conventional control container. Regarding the reagent, the performance of the reagent deteriorates with time, and after 14 days of storage, it shows a value that is about 10% lower than the activity value on the day of storage, indicating that the reagent can no longer be used as a reagent. .

The pH of the first reagent on the storage start day is 7.
On the other hand, after 14 days of storage, the pH of the first reagent stored in the reagent storage container A was 7.97, and the pH of the first reagent stored in the reagent storage container B was 7.98. The pH of the first reagent contained in the control container was 7.93.

Due to the above, in the reagent storage container of the present invention whose internal volume decreases in accordance with the decrease in internal volume, it is possible to completely shut off the reagent contained in the container and the outside air, and therefore the reagent It was confirmed that stable storage could be performed without deteriorating the performance of and the accurate measurement could be performed.

Example 2 Storage stability test of alkaline phosphatase measurement reagent stored in a reagent storage container whose internal volume decreases as the internal volume decreases

Alkaline phosphatase (A) contained in a reagent container whose internal volume decreases as the internal volume decreases
The storage stability of the (LP) measuring reagent was tested.

(1) A reagent consisting of 1.0 M diethanolamine buffer solution (pH 9.8) was prepared and used as the first reagent for measuring alkaline phosphatase (ALP). (2) 1.0 M diethanolamine buffer solution containing 50 mM disodium p-nitrophenyl phosphate (pH
A reagent consisting of 9.8) was prepared and used as a second reagent for measuring alkaline phosphatase (ALP).

(3) The first reagent of (1) and the second reagent of (2) are added to the reagent storage container A of (2) of Example 1, respectively.
Was filled. (4) Reagent storage container A storing the first reagent of (3)
And the reagent storage container A containing the second reagent is placed in a reagent storage of the 736-15 type automatic analyzer manufactured by Hitachi, Ltd. where the temperature is kept at 10 ° C., and each of these tubes is used in the automatic analyzer. It was connected to the reagent switching valve. And saved.

(5) As a control, the control container of (9) of Example 1 was filled with the first reagent of (1) and the second reagent of (2). Next, the control container containing the first reagent and the control container containing the second reagent were stored in the reagent container in which the temperature of the Hitachi 736-15 type automatic analyzer was kept at 10 ° C as in (4). Place it in the container and insert it into the control container through the opening. The other end of a tube made of tetrafluoroethylene-hexafluoropropylene copolymer (FEP) with an inner diameter of 1 mm and a length of 60 cm is connected to the reagent switching valve of this automatic analyzer. Connected to. And saved.

(6) The stability of each of the reagents (4) and (5) stored in a reagent container at 10 ° C. was tested. Using the first reagent and the second reagent stored in the reagent storage container A and the first reagent and the second reagent stored in the control container, respectively, the start date of storage, after 7 days, after 14 days, after 21 days and after 30 days , Alkaline phosphatase activity value 202I
The measurement of U (international unit) / L serum sample was performed in quintuplicate. The measurement with this Hitachi 736-15 type automatic analyzer was carried out by adding 300 μL of the first reagent to 5 μL of the sample and incubating at 37 ° C. for 3 minutes, and then adding 75 μL of the second reagent at 37 ° C. Main wavelength 450 nm and sub wavelength 5
The alkaline phosphatase activity value was calculated from the rate of increase in absorbance from 54 seconds to 4 minutes and 30 seconds after addition of the second reagent at 46 nm.

The test results of the stability of this stored reagent are shown in FIG. In FIG. 2, the horizontal axis represents the number of days elapsed after the start of storage, and the vertical axis represents the relative activity value when the alkaline phosphatase activity values of the five-fold measurement were averaged and divided by the alkaline phosphatase activity value on the storage start day. Represent

[0058]

FIG. 2

From this figure, it can be seen that the reagent stored in the reagent storage container A of the present invention shows the same alkaline phosphatase activity value as the storage start date even after 30 days storage at 10 ° C. However, in the case of the reagent stored in the conventional control container, the performance of the reagent deteriorates with time, and after 30 days of storage, the activity value of the storage start date is decreased. Than 7.3%
Also shows a low value and can no longer be used as a reagent.

The pH of the first reagent on the storage start day is 1
After 30 days of storage, the pH of the first reagent stored in the reagent storage container A was 10.14, and the pH of the first reagent stored in the control container was 10.03. Was becoming.

Due to the above, in the reagent storage container of the present invention whose internal volume decreases in accordance with the decrease in internal volume, it is possible to completely shut off the reagent contained in the container and the outside air, and therefore the reagent It was confirmed that stable storage could be performed without deteriorating the performance of and the accurate measurement could be performed.

Example 3 Storage stability test of alkaline phosphatase assay reagent [reagent according to the Japanese Society for Clinical Chemistry Recommendation] stored in a reagent storage container whose internal volume decreases as the internal volume decreases

Alkaline phosphatase (A) stored in a reagent storage container whose internal volume decreases as the internal volume decreases
The storage stability of the LP) measuring reagent [reagent conforming to the method recommended by the Japan Society for Clinical Chemistry] was tested.

(1) A reagent consisting of 1.0 M 2-ethylaminoethanol buffer (pH 9.9) was prepared and used as the first reagent for measuring alkaline phosphatase (ALP). (2) A reagent consisting of 1.0 M 2-ethylaminoethanol buffer (pH 9.9) containing 75 mM disodium p-nitrophenyl phosphate was prepared and used as a second reagent for alkaline phosphatase (ALP) measurement reagent. did.

(3) The first reagent of (1) and the second reagent of (2) are added to the reagent storage container A of (2) of Example 1, respectively.
Was filled. (4) Reagent storage container A storing the first reagent of (3)
And the reagent storage container A containing the second reagent is placed in a reagent storage of the 736-15 type automatic analyzer manufactured by Hitachi, Ltd. where the temperature is kept at 10 ° C., and each of these tubes is used in the automatic analyzer. It was connected to the reagent switching valve. And saved.

(5) As a control, the control container of (9) of Example 1 was filled with the first reagent of (1) and the second reagent of (2), respectively. Next, the control container containing the first reagent and the control container containing the second reagent were stored in the reagent container in which the temperature of the Hitachi 736-15 type automatic analyzer was kept at 10 ° C as in (4). Place it in the container and insert it into the control container through the opening. The other end of a tube made of tetrafluoroethylene-hexafluoropropylene copolymer (FEP) with an inner diameter of 1 mm and a length of 60 cm is connected to the reagent switching valve of this automatic analyzer. Connected to. And saved.

(6) The stability of each of the reagents (4) and (5) stored in a reagent container at 10 ° C. was tested. Using the first reagent and the second reagent stored in the reagent storage container A and the first reagent and the second reagent stored in the control container, respectively, the start date of storage, after 7 days, after 14 days, after 21 days and after 30 days , Alkaline phosphatase activity value 253I
The measurement of U (international unit) / L serum sample was performed in quintuplicate. The measurement with this Hitachi 736-15 type automatic analyzer was carried out by adding 300 μL of the first reagent to 5 μL of the sample and incubating at 37 ° C. for 3 minutes, and then adding 75 μL of the second reagent at 37 ° C. Main wavelength 415 nm and sub wavelength 5
Second reagent addition at 05 nm 1 min 18 sec 4 min 3
It was performed by calculating the alkaline phosphatase activity value from the rate of increase in absorbance up to 0 seconds.

The test results of the stability of this stored reagent are shown in FIG. In FIG. 3, the horizontal axis represents the number of days elapsed after the start of storage, and the vertical axis represents the relative activity value when the alkaline phosphatase activity values of five-fold measurement were averaged and divided by the alkaline phosphatase activity value on the storage start day. Represent

[0069]

FIG. 3

From this figure, it can be seen that the reagent stored in the reagent storage container A of the present invention shows the same alkaline phosphatase activity value as that on the storage start day even after 30 days storage at 10 ° C. However, in the case of the reagent stored in the conventional control container, the performance of the reagent deteriorates with time, and after 30 days of storage, the activity value of the storage start date is decreased. Than 7.2%
Also shows a low value and can no longer be used as a reagent.

The pH of the first reagent on the storage start day was 1
While it was 0.30, the pH of the first reagent stored in the reagent storage container A after storage for 30 days was 10.30, and the pH of the first reagent stored in the control container was 10.23. Was becoming.

Due to the above, in the reagent storage container of the present invention whose internal volume decreases in accordance with the decrease in internal volume, it is possible to completely shut off the reagent contained in the container and the outside air, and therefore the reagent It was confirmed that stable storage could be performed without deteriorating the performance of and the accurate measurement could be performed.

Example 4 Storage stability test of a zinc sulfate turbidity test measurement reagent stored in a reagent storage container in which the internal volume decreases as the internal volume decreases

The storage stability of the zinc sulfate turbidity test (ZTT) measurement reagent stored in a reagent storage container whose internal volume decreases as the internal volume decreases was tested.

(1) 24 mg / L of zinc sulfate (ZnS)
O ₄ .7H ₂ O), 302 mg / L 5,5′-diethylbarbituric acid and 190 mg / L sodium barbital (pH 7.60) were prepared and used as a zinc sulfate turbidity test (ZTT) measurement reagent. did.

(2) The measurement reagent of (1) was filled in the reagent storage container A of (2) of Example 1. (3) Reagent storage container A containing the measurement reagent of (2)
Was placed in a reagent storage in which the temperature of Hitachi 736-15 type automatic analyzer was kept at 10 ° C., and this tube was connected to the reagent switching valve of this automatic analyzer. And saved.

(4) As a control, the control container of (9) of Example 1 was filled with the measurement reagent of (1). Next, the control container containing this measurement reagent was placed in the reagent container in which the temperature of the Hitachi 736-15 type automatic analyzer was kept at 10 ° C as in (3), and the inside of the control container was opened from the opening. The other end of the inserted tube made of tetrafluoroethylene-hexafluoropropylene copolymer (FEP) having an inner diameter of 1 mm and a length of 60 cm was connected to the reagent switching valve of this automatic analyzer. And saved.

(5) The stability of each of the reagents (3) and (4) stored in a reagent container at 10 ° C. was tested. Using the measurement reagent stored in the reagent storage container A and the measurement reagent stored in the control container, respectively, the start date of storage and after 7 days of storage,
After 14 days, 21 days, and 30 days, a serum sample of 7.8 Kunkel unit (Kunkel unit) was measured in quintuplicate. In addition, in the measurement with this Hitachi 736-15 type automatic analyzer, 350 μl of the measurement reagent was added to 7 μL of the sample.
μL was added and reacted at 37 ° C., and the absorbance at 7 minutes and 30 seconds after the addition of the measuring reagent at the main wavelength of 600 nm and the sub-wavelength of 700 nm was measured. From this absorbance, the absorbance of the reagent blank (reagent blind test) was subtracted from zinc sulfate. The turbidity test was performed by calculating the measured value. As a reference, the measurement reagent of (1) was prepared at each measurement, and the measurement was performed in the same manner as described above.

The test results of the stability of this stored reagent are shown in FIG. In addition, in FIG. 4, the horizontal axis represents the number of days elapsed after the start of storage, and the vertical axis represents the average value of the five-fold measurement zinc sulfate turbidity test measurement value. Represents the relative activity value when divided by the test measurement.

[0080]

FIG. 4

From this figure, in the reagent stored in the reagent storage container A of the present invention, even after 30 days of storage at 10 ° C., the same zinc sulfate turbidity test measurement value as the reagent prepared at the time of measurement was obtained. It can be shown and accurate measurement can be performed. On the other hand, in the reagent stored in the conventional control container, the performance of the reagent deteriorates with time, and after 30 days of storage, It can be seen that the measured value of the reagent prepared at the time of measurement is as high as 12%, and the reagent can no longer be used as a reagent.

Further, the pH of the measurement reagent on the storage start day was 7.
However, after 30 days of storage, the pH of the measurement reagent stored in the reagent storage container A was 7.54, and the pH of the measurement reagent stored in the control container was 7.48.

Due to the above, in the reagent storage container of the present invention whose internal volume decreases in accordance with the decrease in internal volume, the reagent contained in the container and the outside air can be completely shut off, and therefore the reagent It was confirmed that stable storage could be performed without deteriorating the performance of and the accurate measurement could be performed.

[0084]

INDUSTRIAL APPLICABILITY The reagent storage container of the present invention whose internal volume decreases in accordance with the decrease in internal volume can prevent outside air from flowing into the container due to the use of the reagent, and can be stored in the container. The reagent can be used even in an analyzer or the like while always completely blocking the reagent and the outside air.

Therefore, in the reagent storage container of the present invention, carbon dioxide in the air as the outside air is dissolved in the reagent, the pH of the reagent is lowered, and oxygen in the air as the outside air is a constituent component of the reagent. Oxidize, the components of the reagent are concentrated by evaporation of water and microorganisms such as bacteria or mold are mixed in the container, and can be prevented from breeding, It can be stored and used for a long time without deteriorating the performance of the reagent, and is economically effective.

Further, according to the method for measuring a test substance of the present invention, which uses a container for storing a reagent whose inner volume decreases according to the decrease of the inner volume, the measuring reagent is stored and used for a long period of time without deteriorating its performance. This is a method that enables accurate measurement for a long period without any error.

[Brief description of drawings]

FIG. 1 is a graph showing the stability test results of a leucine aminopeptidase measurement reagent stored and stored in a reagent storage container whose internal volume decreases as the internal volume decreases.

FIG. 2 is a graph showing stability test results of an alkaline phosphatase measurement reagent stored and stored in a reagent storage container in which the internal volume decreases as the internal volume decreases.

FIG. 3 is a graph showing the stability test results of an alkaline phosphatase assay reagent [reagent according to the Japanese Society for Clinical Chemistry Recommendation] stored in a reagent storage container whose internal volume decreases as the internal volume decreases. Is.

FIG. 4 is a graph showing the stability test results of a zinc sulfate turbidity test measurement reagent stored and stored in a reagent storage container in which the internal volume decreases as the internal volume decreases.

FIG. 5 is a diagram showing an example of a bag-shaped reagent storage container in which the internal volume decreases as the internal volume decreases.

[Explanation of symbols]

 1 Fusion part 2 Reagent storage part 3 Tube

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Inventor Kazunori Takakura 1-56 Kanda Jinbocho, Chiyoda-ku, Tokyo Shinotest Co., Ltd.

Claims (6)

[Claims] 1. A reagent storage container, the internal volume of which decreases as the internal volume decreases. 2. The reagent storage container according to claim 1, wherein the reagent storage container is a container that is deformable and molded so as to be hermetically sealed. 3. The reagent storage container according to claim 1, wherein the reagent storage container has a partition wall that is movable or deformable inside the container and is molded so as to be hermetically sealed. 4. A method for measuring a test substance using a reagent storage container, the internal volume of which decreases as the internal volume decreases. 5. The method for measuring a test substance according to claim 4, wherein the reagent storage container is a deformable container and is formed so as to be hermetically sealed. 6. The method for measuring a test substance according to claim 4, wherein the reagent storage container is a container that has a partition wall that is movable or deformable inside the container and that is molded so as to be hermetically sealed.