JPH02249970A - Instrument for measuring solution component - Google Patents

Abstract

PURPOSE:To prolong the life of a reagent and calibrating liquid by providing a reagent vessel connected to a detector in a thermostatic chamber which maintains the vessel at a room temp. or below. CONSTITUTION:After a sample liquid S is supplied by a pump 7 to an injector 10, the injector 10 is rotated to switch the flow passage to the flow passage of a carrier liquid C1 and the sample liquid S cut off in such a manner is inserted by the carrier liquid C1 fed by a pump 6 and is mixed 12. The flow passage of the sample liquid S is connected to the flow passage from a calibrat ing liquid vessel 1 by a 3-way valve 5 just before the pump 7. The calibrating liquid can be supplied to the ejector 10 in place of the sample liquid S by switching the 3-way valve 5 at the time of calibration. On the other hand, the reagent is supplied from the reagent vessel 2 by a pump 9 to another injector 11 and the reagent cut off by a similarly switching the injector 11 is inserted by the carrier liquid C2 and is mixed 13 by a pump 8. The liquid mixture from the mixers 12, 13 is further mixed 14 and is passed through a detector 15, from which the liquid mixture is discharged. This liquid mixture is processed 16 as an electrical signal and the concn. thereof is determined.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an apparatus for measuring the concentration of solution components using a flow injection analysis method or the like.

[Conventional technology]

Various analytical devices have been developed that perform conventional wet analysis using flow injection analysis and the like.

For example, a solution component measurement device that uses flow injection analysis uses an injector with a switchable flow path to collect sample liquid and reagent separately, and then switches the flow path of the injector to the reactor, measurement cell, or detector. The components to be measured in the sample solution can be detected by supplying the sample solution to the flow path of the sample solution and mixing them, and detecting the product of the reaction between the sample solution and the reagent using an electrode, etc., or by detecting the amount of luminescence accompanying the reaction. It automatically measures concentration and is already used mainly in the medical field.

In measurement devices such as flow injection analyzers, if the reaction or detection used for measurement is affected by temperature, the reactor, measurement cell, or detector built into the device should be kept at a temperature above room temperature suitable for the reaction or detection. By keeping the temperature constant, the influence of temperature on reaction and detection is avoided.

However, in conventional solution component measuring devices, including flow injection analysis Ml, reagents and calibration solutions used for reactions are placed near the device in glass or plastic containers, so they cannot be left at room temperature. there were.

For this reason, in analyzes that use thermally unstable reagents and calibration solutions, including enzyme-based reagents, these reagents and calibration solutions are susceptible to deterioration due to elevated ambient temperatures in heated rooms, not only in summer but also in winter. However, the maintenance cycle is extremely short, as the activity rapidly deteriorates and the service life reaches the end within a short period of time, requiring replacement.

In addition, thermally unstable reagents and calibration solutions such as enzyme-based reagents
If left at room temperature, the lifespan will be extremely shortened, so it has been impossible to incorporate reagent and calibration solution containers into a measuring device such as a 70-injection analyzer.

[Problem to be solved by the invention]

In view of such conventional circumstances, the present invention is a solution component that can extend the life of thermally unstable reagents and calibration solutions, and can also be used with reagents and calibration solutions that were previously unusable in terms of lifespan or durability. The purpose is to provide a measuring device for.

[Means to solve the problem]

In order to achieve the above object, the present invention provides an apparatus for feeding a sample liquid and a reagent to a detector and measuring the concentration of a component to be measured in the sample liquid using the reaction between the sample liquid and the reagent. , the reagent container connected to the detector via a conduit is provided in a constant temperature bath capable of maintaining the reagent at a temperature below room temperature suitable for storage.

Furthermore, if calibration is also performed automatically, the reagent containers and calibration solution containers connected to the detector via piping must be identical and capable of maintaining the reagents and calibration solutions at a humidity below room temperature suitable for storage. Or, it is characterized by being provided in a separate constant temperature bath.

[Effect]

In the measuring device of the present invention, the reagents and/or calibration solutions stored in the container can be maintained at a temperature below room temperature suitable for storage, so not only during measurement but also when not measuring (including when not in use). Deterioration of reagents and calibration solutions can be prevented. As a result, the life of the device can be dramatically extended while retaining thermally unstable reagents and calibration solutions within the device.

For example, in the analysis of adenosine triphosphate (ATP), ATP is decomposed into adenosine monophosphate (AMP) by the catalytic action of luciferase in the presence of luciferin and Mg2+, and the amount of luminescence accompanying the following reaction is determined. Measure the ATP concentration from: AMP+orluciferin+Co+ppi+hν
This analytical method has high specificity for the above reaction, and emission spectrometry is an effective analytical method with a wide dynamic range.
Since the luciferin and luciferase used are thermally unstable and have durability problems, it has been difficult to convert them into measuring devices using conventional flow injection methods.

However, according to the present invention, the luminescent reagent luciferin
Since the life of the luciferase solution etc. can be dramatically extended, it is possible to construct a measuring device incorporating the above reagent.

Furthermore, even if there are reagents that have better thermal stability than luciferin or luciferase, there are many that are preferable to be used at a temperature lower than room temperature than at room temperature. With the measuring device used, it is also possible to significantly extend the maintenance period for replacing reagents, etc.

Note that the degree of deterioration due to temperature varies depending on the reagent and calibration solution used, so it is preferable to set the temperature setting of the constant temperature bath containing the reagent container and calibration solution container depending on the reagent and calibration solution used. The temperature is usually about 4σ or lower. However, it is of course necessary to avoid temperatures so low that the reagents and calibration solutions freeze.

As such a constant temperature bath, for example, a constant temperature bath having a cooling function and whose temperature can be adjusted can be suitably used.

〔Example〕

FIG. 1 shows a specific example of the device of the present invention, which is a measuring device for automatically analyzing ATP using a flow injection analysis method.

The calibration solution container 1 is filled with an ATP calibration solution, and the reagent container 2 is filled with a luciferin/luciferase solution containing luciferase, luciferin, and Mg2+, and these are stored separately in temperature-adjustable thermostats 3 and 4, respectively. .

After the sample liquid S containing ATP is supplied to the injector 10 by the pump 7, the injector 10 is rotated to fill the flow path with carrier liquid 0 (HEPIC8 with pH 7.75).
By switching to the flow path of the buffer solution), the cut sample solution S is transferred to the carrier solution C fed by the pump 6.
It is sandwiched between the two and sent to the mixer 12 where it is mixed. In addition, the flow path of the sample liquid S is connected to the flow path from the calibration liquid container 1 by a three-way valve 5 before the pump 7, and during calibration, the three-way valve 5 is switched and the calibration liquid is injected into the injector 10 instead of the sample liquid S.
It is now possible to supply

On the other hand, the reagent is supplied from the reagent container 2 to another injector 11 by the pump 9, and by switching the injector 11 in the same way as above, the cut out reagent is sandwiched in the carrier liquid C (carrier liquid C is the same) and pumped. 8 to a mixer 13 for mixing.

The mixed liquid containing the sample liquid S sent out from the mixer 12 and the mixed liquid containing the reagent sent out from the mixer 13 are combined, further mixed in the mixer 14, passed through the detector 15, and discharged. Ru. In the detector 15, the amount of luminescence accompanying the reaction is detected by a chemiluminescence detector or the like, and is sent as an electrical signal to the processing device 16, where the data is processed and the ATP concentration is determined.

In this automatic measuring device, the temperature of the constant temperature bath 3 containing the calibration solution container 1 is set to 4σ (condition 1), room temperature (condition 2) and 30℃.
C” (condition 3) and 10”m under each of the above conditions.
Changes in the residual activity of the 0 l/l ATP calibration solution over time were measured. In addition, the residual activity of the ATP calibration solution is
The output of the detector 15 obtained by sucking 10 mol IATP solution adjusted each time for measurement from the flow path of the sample liquid S, and the detection obtained with the ATP calibration solution sucked from the calibration solution container 1 by switching the three-way valve 5. It was determined from the ratio with the output of the device 15. Furthermore, the luciferin/luciferase solution used was adjusted to a constant concentration each time the measurement was performed.

As can be seen from the results shown in Figure 2, when comparing the elapsed time when the residual activity of ATP reaches 1/2 as a guideline for lifespan, it is only 9 days under condition 3 and 16 days under condition 2. Therefore, under condition 1 according to the present invention, the lifespan was more than doubled to 37 days.

Similarly, the temperature of the constant temperature bath 4 containing the reagent container 2 was set to 4C.
(Condition 1), room temperature (Condition 2), and 30C (Condition 3).
Changes in the residual activity of the luciferase solution were measured.
U/@/ ), luciferin (2X 10= mol
/l) and Mg'' (10 mol/l).The ATP solution in this case was adjusted for each measurement.
A concentration of 0-6 mol/l was used.

The results are shown in Figure 3. Comparing the elapsed time when the remaining lifespan of the luciferin-luciferase solution becomes 172 as a lifespan guideline, it is only 1 day in condition 3 and 4 days in condition 2, whereas it is only 10 days in condition 1 according to the present invention. It can be seen that the lifespan is dramatically extended by exceeding the 4σ value and maintaining it at 4σ.

When the apparatus was not in use, such as at night, the calibration solution container 1 and reagent container 2 were removed from the apparatus and stored at an appropriate temperature in a refrigerator.

〔Effect of the invention〕

According to the present invention, the lifespan of reagents and calibration solutions used in a solution component measuring device can be dramatically extended, and a measuring device that uses reagents that were conventionally considered to have a short lifespan and was difficult to develop into a device. The range of substances to be measured can be expanded.

At the same time, even if the reagents are conventionally used as devices, the maintenance cycle of the measuring device can be significantly lengthened by extending their lifespan.

Furthermore, reagents such as luciferin and luciferase are often extremely expensive, but by extending their lifetime, they are extremely advantageous economically.

[Brief explanation of drawings]

FIG. 1 is a schematic configuration diagram showing a specific example of the present invention. Figure 2 is a graph showing the relationship between residual activity and elapsed time depending on the holding temperature of the ATP calibration solution, and Figure 3 is a graph showing the relationship between residual activity and elapsed time depending on the holding temperature of the luciferin/luciferase solution. It is. 1. Calibration liquid container 2. Reagent container 3.4. Constant temperature bath 5. Three-way valve 6.7.8.9. Pump 10.11. Injector 12.13.14. Mixer 15.・Detector 16...Processing equipment procedure correction equipment (voluntary) Elapsed time (days) on May 25, 1990 1゜Indication of incident Name of Heisei invention Year Patent Application No. 71435 Solution component measuring device No. Correction Relationship with cases involving persons who commit crimes

Claims (2)

[Claims] (1) In a device that sends a sample liquid and a reagent to a detector and measures the concentration of a component to be measured in the sample liquid using the reaction between the sample liquid and the reagent, the sample liquid and reagent are sent to the detector via a pipe line. 1. An apparatus for measuring solution components, characterized in that a connected reagent container is provided in a constant temperature bath capable of maintaining the reagent at a temperature below room temperature suitable for storage. (2) A device that sends a sample solution or calibration solution and a reagent to a detector and measures the concentration of the component to be measured in the sample solution or calibration solution using the reaction between the sample solution or calibration solution and the reagent. In this method, a reagent container and a calibration solution container connected to a detector via a conduit are provided in the same or separate thermostatic chambers capable of maintaining the reagent and calibration solution at a temperature below room temperature suitable for storage. A device for measuring solution components, characterized in that: