JPH03214056A - Automatic analyzing apparatus - Google Patents

Abstract

PURPOSE:To maintain analyzing accuracy by computing the rate of dilution of a sample based on the measured results of sample diluent itself and the internal standard material in the diluted mixed liquid of the sample and the diluent, and obtaining the rate of dilution of the sample at high accuracy. CONSTITUTION:A control device 20 compares the amount of the liquid of a sample with the amount of the sample required for performing the analysis of items to be analyzed. When the sufficient amount of the sample is present, the original sample is used and analyzed. Meanwhile, when the sample in a sample container 6 is less than the amount of the sample required for the analysis, the device 20 prepares the required amount of the mixed liquid of the sample and diluent. When each reaction container 2 containing the specified amounts of the sample and the mixed liquid of the sample and the diluent is moved to a reagent distributing mechanism 14, the device 20 distributes the specified amounts of the reagent suitable for detecting the items of analyses and the reagent suitable for detecting the internal standard material contained in the sample diluent into the each container 2. When each container 2 reaches a light measuring device 16, the absorbance of the intended component and the absorbance of the internal standard material contained in each container 2 are measured. Thus the rate of dilution of the original sample is obtained. The concentration of the intended component is obtained based on a specified operation expression.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention relates to an automatic analyzer suitable for analyzing a minute amount of sample.

(Prior art) Automatic analyzers usually dispense a specified amount of the sample collected in a sample container into a reaction chamber, mix it with a reagent suitable for horizontally comparing the target components, and then use a photometric device. The device is configured to be transferred throughout the body. Such an automatic analyzer is usually configured to dispense a minute amount of a sample from a sample container into a reaction container using a sample dispensing pipettor nozzle in the amount of several to several tens of microliters per analysis item.

By the way, since the tip of such a sample dispensing nozzle is extremely delicate, its position is normally controlled to ensure a certain gap 1jj between it and the bottom of the sample container. When analysis is completed, at least 100 to several hundred microliters of sample remains in the sample container, which limits the amount of sample that can be collected.When analyzing body fluids from newborns or small experimental animals, automatic analysis is recommended. The sample remaining at the bottom of the sample container of the device is taken out and used for manual analysis.

(Problem to be solved by the invention) According to this, while it is possible to use samples effectively, the number of samples required for analysis per one analysis item is several times that when using automatic analyzer M. The amount increases by about 20 times, which not only reduces sample utilization efficiency, but also makes analysis work time-consuming.

In order to solve this problem, the amount of sample is increased by measuring a sample diluent to the desired dilution ratio and mixing it with the original sample. Since measurement involves extremely large errors, there is a problem in that analysis results include large errors.

The present invention has been made in view of these problems, and its purpose is to provide a novel method that can maintain analysis accuracy by accurately grasping the dilution rate of a sample whose molar volume has been increased by dilution. Automatic analyzer M! i! It is about providing.

(Means for Solving the Problem) In order to solve such problems, the present invention provides a sample diluent container containing a sample diluent containing an internal standard substance, and a sample diluent container containing a sample diluent containing a sample and the sample diluent. A means for aspirating and dispensing a predetermined amount of sample diluent into a reaction container, a means for measuring an internal standard substance in the reaction container, and a means for measuring an internal standard substance in the sample diluent itself and the diluted sample dilution mixture. A means is provided for calculating the dilution rate of the sample derived from the results and calculating the concentration of the analytical component in the original sample.

(Function) When the original sample is insufficient, a diluent is added and diluted to the original sample to prepare a sample dilution mixture and dispensed into a reaction container. This allows you to increase the amount of sample and find out the dilution rate of the sample from the concentration of the internal standard contained in it and the measurement results of the internal standard in the original sample diluent.
Therefore, there is no need to carry out measuring operations for minute quantities. For this reason,
Error factors caused by measurement errors are eliminated, and minute amounts of samples can be analyzed with high precision.

(Example) The details of the present invention will be described below based on illustrated examples.

FIG. 1 shows an embodiment of the present invention, and reference numeral 1 in the figure is a transport mechanism that circulates and moves reaction vessels 2, 2, 2, etc., and is the starting point of the analysis work. A sampling mechanism 3, which will be described later, is also provided. This sampling mechanism 3 is moved by a robot arm 4 between a sample container 6 of a sample container mount 5, a sample diluent storage tank 8, which will be described later, and a reaction container 2, and moves between the sample container 6 and the sample dilution liquid storage tank. 8, a sample dispensing nozzle 7 that descends into the reaction vessel 2, and a plunger pump 9.

FIG. 2 shows an embodiment of the sample dispensing mechanism, and reference numeral 11 in the figure is a liquid level detection electrode arranged along the bottom nozzle 7 for dispensing the sample. It is configured to output a signal when it comes into contact with the liquid surface together with the ivy nozzle 7.

Returning again to FIG. 1, the reference numeral 14 in the figure is a reagent dispensing mechanism, and the nozzle 2 is movable between the reagent stand 15 and the reaction container 2.
2, and is configured to aspirate the reagent passed through the analysis item and a reagent suitable for detecting an internal standard substance from the reagent holder 15 and dispense it into the reaction container 2 through the nozzle 22.

Reference numeral 16 denotes a photometer, which includes a flow cell 17 that accommodates a mixed reaction solution of a sample and a reagent from the reaction container 2, and a light emitting element 18 that measures the absorbance of the mixed reaction solution sucked into the flow cell 17.
, and a light receiving element 19.

20 is a control device consisting of a microcomputer that controls analysis operations, and includes a liquid level detection electrode 11 and a photometric device N1.
6 is input, and is configured to carry out the operations shown in the flowchart described later. Note that the reference numeral 10 in the figure indicates a stirring device W for uniformly mixing the solution in the reaction container 2.

Next, the operation of the apparatus configured as described above will be explained based on the flowchart shown in FIG.

Prior to analysis work, use succinic acid, oxalic acid, and citric acid when analyzing time-stable and inexpensive substances, such as serum, without affecting the analytical components to be analyzed. , various organic acids such as α-ketoglutaric acid, β-hydroxybutyric acid, biruvic acid, etc., as well as fructose, sucrose, etc.! II! , L-glutamate, etc., dissolved approximately 100 times as much as the highest expected concentration in the sample, is stored in the sample diluent storage tank 8 as a diluent.

Prior to analysis, the control device 20 aspirates a predetermined amount of the sample diluent in the sample diluent storage tank 8 through the bivet nozzle 7 and dispenses it into the reaction container 2 (Step 1), and the reaction container 2 is placed in parallel with the analysis work. or preferentially transfer it to the reagent dispensing machine 1J1114, and dispense the reagent for internal standard detection using the reagent dispensing nozzle 22 (step
), the control device i20 converts this reaction container 2 into a photometric bag! [
16, measure the internal standard substance contained in the sample diluent itself, and calculate its absorbance! Memorize δ (step Ha).

Next, the control table! Step 20 is step 2) of moving the throat nozzle 7 of the sample dispensing mechanism 3 to a predetermined position and setting a counter (not shown).
The sample dispensing nozzle 7 is lowered toward the sample container 6, and at the same time the counter is caused to perform a counting operation (Step E). When the liquid level detection electrode 11 comes into contact with the sample due to the lowering of the sample dispensing nozzle 7, the liquid level A signal is output from the detection electrode 11 (to step 2), and the control device 20 calculates the total amount of the original sample liquid from the counted value (step 2).

That is, the count value N (second
(Figure) represents the liquid level of the sample, that is, the total amount of the original sample.

The control device f20 compares the liquid level of the sample with the amount of sample required to execute the preset analysis item (step H), and if there is a sufficient amount of the sample, the amount of sample is Analyze the specified analysis items using the original sample without diluting the sample (step re).

On the other hand, if the number of samples in the sample container 6 is smaller than the number of samples required for processing the preset type of analysis,
The control table W20 moves the dowel nozzle 7 to the sample dilution liquid storage tank 8, and injects a predetermined amount of sample dilution liquid in order to prepare the amount of sample dilution mixture liquid necessary to process a preset type of analysis. to be absorbed. Next, the sample pipettor nozzle 7 is moved to the sample container 6, and the sample dilution liquid is discharged into the sample container 6 (Step N). In this process, the original sample and the sample dilution liquid are stirred and mixed uniformly. However, if more even stirring is required, the original sample and diluted sample solution can be sucked into the dot nozzle and moved up and down, and the stirring and mixing can be repeated by repeating suction and discharge. The sample diluted mixture prepared in this manner is then dispensed to analyze the preset analysis items and the concentration of the internal standard substance (stepping).

The control table M20 indicates that when each reaction container 2 containing a predetermined amount of sample and sample dilution mixture is moved to the reagent dispensing mechanism 14, a reagent suitable for detecting an analysis item and a sample dilution liquid are added. A predetermined amount of a reagent suitable for detecting the internal standard substance contained therein is dispensed into each reaction vessel 2 (Step O), and then when each reaction vessel 2 reaches the photometer 16, each The absorbance A of the target component and the absorbance Al80 of the internal standard substance contained in the reaction chamber 2 are measured (step wa). be measured.

As a result, the absorbance AI of the internal standard substance in the sample dilution mixture just measured! . and the ratio (IA
+s. XA,s) will represent the dilution rate of the original sample (step force).

Therefore, the concentration CA of the target component can be obtained by performing calculations based on the following calculation formula (step YO).

In other words, if the amount of sample dispensed into the reaction container 2 is different between the time of sample dilution mixture analysis and the time of urine sample analysis, Cs = (A^-ARX (vo/v)X (V/V)
o)) XKX (v/vo)X (Vo/V)X
A +s/(A +s A +go)= (AAx
(v/vo)X (Vo/V) -A))X to x A
+s/(A +s A +so) ・・・(
1) However, AA is the reaction absorbance Am of the diluted sample mixture of analytical component A, and the absorbance of the reagent blank solution for analytical component A under the original sample analysis conditions is the concentration conversion factor of analytical component A under the original sample analysis conditions. V is the amount of original sample sampled when analyzing component A. is the sampling amount of the sample dilution mixture during analysis to the analytical component vo is the total reaction volume during analysis of analytical component A in the sample dilution mixture ■ is the total reaction to the analytical component in the original sample during analysis Each represents the amount of liquid.

In addition, when analyzing the original sample and sample diluted mixture using the same sample volume, (v/vo)X (Vo, /V) =
1, and the tCC tube blank liquid absorbance is the same when analyzing the diluted sample mixture and when analyzing the original sample, so the reciprocal of the dilution rate of the original sample (I A + so / A + s) is
(A + s A + so ) / A + to )
A +s/(A +s A +go)...(2
), the concentration of the target component can be determined.

Thereafter, such a process is repeated to analyze the necessary analytical components of each sample.

Needless to say, in the above process, there is no step to measure the absolute amount of the original sample amount and the amount of the sample dilution mixture when calculating the dilution rate, so there is a large measurement error that tends to occur when measuring minute amounts. Without that, therefore positive? i [
The dilution ratio can be calculated.

In this example, the concentration is corrected all at once after the end of the measurement, but it is clear that the same effect can be obtained even if the concentration is corrected one by one after the analysis of each analytical component is completed. It is.

In addition, in this example, the internal standard of the sample dilution solution was used at the beginning of the analysis. 1! Although the concentration of substance #j is measured and used from now on, it is also possible to measure it for each sample or every certain period of time and update the memory data.

Furthermore, in this embodiment, one correction operation 11i is executed every time the analysis of each analytical component is completed, but for example, when all the components of one sample are measured, It is clear that one step may be taken to perform a correction calculation.

(Effects of the Invention) As explained above, in the present invention, there is provided a sample diluent container containing a sample diluent containing an internal standard substance, and a predetermined amount of the sample diluent prepared from the sample and the sample diluent. A means for aspirating M and dispensing it into a reaction container, a means for measuring an internal standard substance in the reaction container, and a method derived from the measurement results of the internal standard substance of the sample dilution itself and the diluted sample dilution mixture. Since we are equipped with a means to calculate the source of analyte i in the original sample by calculating the dilution rate of the sample, even if the amount of the original sample is small, it is possible to increase the amount by dilution and analyze the required analyte. In addition to making it processable, since the dilution rate is calculated based on the measurement results of the internal standard substance, it is possible to reduce measurement errors as much as possible and maintain a high analytical edge/i.

Furthermore, since the amount of original sample required for analysis can be reduced as much as possible, it is also possible to observe temporal changes in objects that are subject to restrictions on sample collection.

[Brief explanation of drawings]

Figure M1 is a configuration diagram of an apparatus showing an embodiment of the present invention, Figure 2 is a diagram showing an embodiment of a sample dispensing nozzle used in the above-mentioned device, and Figure 3 is a diagram showing the operation of the above-mentioned apparatus. FIG. 1...Transportation mechanism 2...Reaction container 3...
・・Sample sampling mechanism 5 ・・・Sample container holder 6 ・・・Sample container 7 ・・・Sample dispensing nozzle 8 ・・・Sample diluent storage tank 14 ・・・Reagent Dispensing mechanism 15... Trial JIIti stand 16... Photometering device Φ

Claims (1)

[Claims] a sample diluent container containing a sample diluent containing an internal standard; a means for aspirating a predetermined amount of the sample diluent prepared from the sample and the sample diluent and dispensing it into a reaction container; A means for measuring the standard substance, and a means for calculating the concentration of the analyte in the original sample by calculating the dilution rate of the sample derived from the measurement results of the internal standard substance of the sample dilution liquid itself and the diluted sample dilution mixture. An automatic analyzer equipped with