JPH1183869A - Quantitative collection device for specimen - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a quantitative collection device for specimens which can accurately and simply collect the specimens. SOLUTION: This device 20 is composed of a cylindrical body 21, a specimen extracting nozzle 23 comprising a piston body 22 which is contained in it and movable fluid-tightly in the cylindrical body 21 and a container 32 in which a protuberance 31 with the same diameter as that of the piston body 22 is raised. Since surplus specimen adhering to an inner surface of the cylindrical body 21 can be removed by inserting the protuberance 31 in the container 32 into the cylindrical body 21, after the specimens are taken in the cylindrical body 21 by inserting the specimens extracting nozzle 23 in the specimen and moving the piston body 22, the specimens can be accurately and simply collected.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a sample quantification and sampling apparatus used for an automatic analyzer and the like.

[0002]

2. Description of the Related Art In automatic analysis in various fields, it is required to quantitatively collect a measurement sample. For example, collection of a sample in liquid chromatography analysis, which is a typical automatic analysis method, and introduction of a sample into a liquid chromatograph are generally performed by an automatic method using an autosampler. FIG. 7 shows a system configuration of a liquid chromatograph including this autosampler. The eluent 5 is sent by the sending pump 4 and passes through the port ba of the six-way valve 7.
The liquid is led to the waste liquid container 1 via the separation column 3 and the detector 2.
On the other hand, a predetermined amount of the sample 12 is suctioned and collected from the sample suction nozzle 11 by the suction pump 10. The sample collected by suction is introduced into the sample loop 8 through the ports dc of the six-way valve 7, and the sample overflowing the sample loop 8 is guided to the waste liquid container 9.

At the time of sample introduction, the six-way valve 7 is switched, and the ports af, ed, and bc are connected to the six-way valve 7 as shown by broken lines, so that the inside of the sample loop 8 is The sample is led to the separation column 3. Note that the portion surrounded by the broken line in FIG. 7 corresponds to the autosampler 6.

In collecting and introducing such a sample, it may be necessary to dilute the sample. In such a case, the sample 12 sampled by suction is discharged into a separately provided dilution tank 13, diluted in the dilution tank 13, and then the diluted sample is introduced into the sample loop 8 in the same manner as described above. .

[0005] The sample suction nozzle 11 used for the above-mentioned sample collection is formed of a hollow cylindrical body as shown in FIG. 8, and the sample suction nozzle 11 sucks and samples the sample 12 by the suction pump 10. However, at the time of sampling, since an extra sample adheres around the sample suction nozzle, accurate sampling is extremely difficult when sampling a sample with high viscosity such as blood or when sampling a small amount of sample. For this reason, there is a problem that the concentration of the diluted sample is not constant, which causes a measurement error in liquid chromatography analysis.

[0006] For this reason, in the case of a sample that requires dilution, Japanese Patent Application Laid-Open No. Hei 4-70564 discloses that the concentration of a blood sample is measured by measuring the absorbance, and the dilution rate is adjusted according to the concentration to obtain a constant value. A method for obtaining a diluted blood sample at a concentration is disclosed. However, this method includes a dilution operation and is limited to a case where the concentration can be measured by absorbance, and it is not easy to improve an apparatus for attaching an apparatus for measuring absorbance to an existing liquid chromatograph apparatus. There is a problem.

Further, a rubber lid is attached to the sample container, and the sample is sucked and collected by penetrating the lid with a sample suction nozzle. It is also practiced to remove it. However, also in this case, the wiping effect becomes insufficient with a high-viscosity sample, and a problem remains in reproducibility.

[0008] Not only in the case of including the above-mentioned dilution operation in liquid chromatography, but also in general analysis, it is required to accurately and simply collect a sample from a sample stock solution.

[0009]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and it is an object of the present invention to provide a sample quantifying and collecting apparatus capable of collecting a sample accurately and easily.

[0010]

According to the present invention, there is provided a sample-quantifying and sampling apparatus comprising: a cylindrical body; and a piston-like body housed in the cylindrical body and capable of reciprocating in a liquid-tight manner within the cylindrical body. , And a projection having the same diameter as the piston-like body is constituted by a container which is provided upright inside.

[0011]

After inserting the sample suction nozzle into the sample and moving the piston-like body to take in the sample into the cylindrical body, the protrusion having the same diameter as the piston-like body is placed inside the container in which the projection is provided. Since the projection can be inserted into the cylindrical body of the sample suction nozzle to remove an extra sample attached to the wall of the cylindrical body, the sample can be accurately and simply collected.

[0012]

DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 is a schematic explanatory view showing an example of a sample quantification and sampling apparatus according to the present invention. The sample quantitative sampling device 20 includes a sample suction nozzle 23 including a cylindrical body 21 and a piston-like body 22 housed therein and capable of reciprocating in a liquid-tight manner within the cylindrical body 21; 22 and a container 32 having the same diameter as the projection 31 erected therein. In FIG. 1, the container 32 is particularly schematically illustrated. The piston-like body 22 can be reciprocated in the tubular body 21 by a driving unit 24. A sample suction port 25 is provided on the side wall of the cylindrical body 21.

The sample quantifying apparatus 20 is used as follows. As shown in FIG. 2A, in a state where the piston-like body 22 is located at the lowermost end of the tubular body 21, the sample quantitative collection device 20
Is moved to the upper part of the sample 12. The sample quantifying device 20 is lowered and inserted into the sample 12. Thereafter, the piston-like body 22 is pulled up by the drive unit 24, so that a fixed amount of the sample 12 is collected in the cylindrical body 21 (FIG. 2 (b)).

The sample quantitative collection device 20 moves to the upper part of the container 32 on which the projection 31 is erected (FIG. 2C). Next, the sample quantitative collection device 20 descends, and the projection 31 fits into the cylindrical body 21. At the same time, the piston-like body 22 is pulled up at the same speed. At this time, an extra sample is removed (FIG. 2 (d)). The space 26 between the protrusion 31 in the cylindrical body 21 and the gap between the piston-like body 22 finally determines the sample amount.

The piston-like body 22 is further raised,
For example, a pipe from the suction pump 10 is attached to the sample suction port 25, and the sample 12 is sucked by the suction pump 10. The sucked sample 12 is sent to an analyzer such as a liquid chromatograph, for example.

When the sample quantifying device 20 is used continuously, for example, as shown in FIG. 3, the sample quantifying device 20 is connected to a container in which the cleaning liquid is stored via the suction pump 10 and the three-way valve 41. 42. The other connection port of the three-way valve 41 is connected to, for example, an analyzer such as a liquid chromatograph. After such connection, the three-way valve 41 is opened in the direction of the container 42, and the cleaning liquid 421 is supplied to the three-way valve 41-the suction pump 10-.
The liquid flows through the sample suction port 25 into the space 26. At the same time, the projection 31 is pulled out from the inside of the cylindrical body 21 so as to raise the sample quantitative collection device 20. Thereby, the inside of the sample suction nozzle 23 and the inside of the container 32 are cleaned. The waste liquid after washing is discharged from a drain port 33 provided in the container 32.

As the sample suction nozzle 23 in the above-described sample quantifying and collecting apparatus 20, a known stainless steel or a stainless steel coated with a resin, ceramic or the like is used.

As described above, the inner wall of the cylindrical body 21 and the piston-like body 22 are in liquid-tight contact with each other. In the above example, the piston-like body 22 is moved up and down by the drive unit 24, but may be fixed vertically without the drive unit 24. In this case, the above operation is performed by vertically moving only the tubular body 21. Further, the projection 31 has the same diameter as the piston-like body 22, but its shape is not particularly limited as long as it can perform the above-described function.

The container 32 in the sample quantification and sampling apparatus 20 of the present invention can also serve as a dilution tank or a reaction tank.
Next, an application example of diluting a sample in liquid chromatography using the sample quantitative collection device 20 will be described.

FIG. 4 shows a configuration example of the apparatus in this case.
The three-way valve 43, the suction pump 10, and the three-way valve 44 are connected to the sample suction port 25 of the sample quantitative collection device 20 through a pipe.
Are connected in series, the three-way valve 44 is connected to the six-way valve 7 and the three-way valve 45 of the liquid chromatograph, and the three-way valve 45 is connected to the container 46 containing the cleaning liquid 461 and the container 47 containing the diluting liquid 471. ing. Also,
A pipe 431 to the container 32 is further connected to the three-way valve 43.

Dilution is performed as follows using the apparatus having the above configuration. First, in the same manner as in FIG.
In the state where the cleaning liquid 461 is collected and measured in the space 26 shown in (d), the cleaning liquid 461 is supplied to the container 3 via the three-way valve 45-the three-way valve 44-the suction pump 10-the three-way valve 43-the pipe 431.
2 and the excess sample in the container 32 is washed away. The wastewater is discarded from the outlet 33. Next, as shown in FIG.
Is pulled up until its lower end is located immediately above the sample suction port 25. Next, as shown in FIG.
The diluent 471 is discharged from the sample suction port 25 through the three-way valve 45-three-way valve 44-suction pump 10-three-way valve 43 while the sample 12 is diluted, and the collected and weighed sample is placed in the container 32. Release into.

Further, a predetermined amount of diluent 471 is added,
Dilution is performed (FIG. 5 (c)). Next, the piston-like body 22 is pushed down, and the diluent 471 remaining in the tubular body 21 is pushed out into the container 32 (FIG. 5 (d)). A fixed amount of the diluted sample is led to the six-way valve 7 of the liquid chromatograph via the pipe 431-three-way valve 43-suction pump 10-three-way valve 44 to perform liquid chromatography measurement. The remaining diluted sample is discharged from the drain port 33 of the container 32, and washing is performed as necessary.

The cleaning liquid 461 and the diluting liquid 471
May be the same composition. If a reaction reagent is used instead of the diluent 471, automatic pretreatment of a sample by a chemical reaction can be performed. In addition, the container 32 includes components (eg, for example) for efficiently performing dilution, washing, pretreatment, and the like.
A heating device, an ultrasonic oscillator, an absorbance measuring device, etc.).

The above-described sample quantifying apparatus 2 of the present invention 2
At 0, the sample suction port 25 is provided in the cylindrical body 21, but it is not essential that the sample suction port 25 is provided at this position. FIG. 6 shows an example in which a sample suction mechanism is separately provided. The mechanism in FIG. 6 is the same as the mechanism in FIG. 4 except that the sample suction port 25 and the pipe from the sample suction port 25 to the three-way valve 43 are not provided. This example is characterized in that the pipe 431 also serves as a sample suction port. Less than,
This mechanism will be described.

First, in the state where the sample 12 is collected and measured in the same manner as in FIG. 2D, the cleaning liquid 461 is supplied to the container via the three-way valve 45-three-way valve 44-suction pump 10-three-way valve 43-piping 431. Into container 32
Wash off excess sample in 2. The wastewater is discarded from the outlet 33. Next, the piston-like body 22 is pushed down while the sample suction nozzle 23 is pulled up, and the collected and weighed sample is discharged into the container 32. Further, the diluent 471 is discharged into the container 32 via the three-way valve 45-three-way valve 44-suction pump 10-three-way valve 43-piping 431. Thus, the sample is diluted with the diluent 471 in the container 32.

The sample suction nozzle 23 is lowered and the cylindrical body 21
Is inserted into the diluted sample in the container 32, the piston-like body 22 is pulled up, and the diluted sample accumulated in the container 32 is sucked into the cylindrical body 21. The sample suction nozzle 23 is raised again, and at the same time, the piston-like body 22 is pushed down, and the diluted sample in the tubular body 21 is discharged into the container 32. By repeating this operation and the above, the cleaning effect of the sample collected in the cylindrical body 21 by the above operation is further enhanced. A fixed amount of the diluted sample in the container 32 and a fixed amount of the diluted sample are led to the hexagonal valve 7 of the liquid chromatograph via the pipe 431-three-way valve 43-suction pump 10-three-way valve 44, and the liquid Perform chromatographic measurements.

[0027]

The present invention will be described in detail below with reference to examples.

Example 1 and Comparative Example 1 The liquid chromatography system shown in FIG. 7 was assembled using the following devices as main devices, and liquid chromatography was performed. (Liquid chromatograph system) Liquid sending pump 4: LC-9A separation column 3: Shimadzu Corporation, Sekisui Chemical Co., Ltd., Micronex
A1c HSII (column using cation exchange resin as a filler) Detector 2: SPD-6AV Autosampler 6 by Shimadzu Corporation: ASU-42 by Sekisui Chemical Co., Ltd.
0. In the first embodiment, the sample circuit portion (specifically, the circuit portion including the suction pump 10, the sample suction nozzle 11, the sample 12, and the dilution tank 13) connected to the port d of the six-way valve 7 of the autosampler 6 is The one modified to the structure shown in FIG. 4 was used, and the one not modified in Comparative Example 1 (FIG. 7)
Having the structure of). Data processing device: Shimadzu Corporation, C-R4A

(Measurement Sample) As a measurement sample, blood collected from a healthy person using sodium fluoride as a blood anticoagulant was used at a concentration of 0.1% by weight of Triton X-10.
A hemolyzed and diluted 100-fold with a phosphate buffer (pH 7) containing 0 was used. (Cleaning liquid) Triton X-100 was used as the cleaning liquid.
(Alkyl phenol ether, manufactured by Tokyo Chemical Industry Co., Ltd.) in 10 mM phosphate buffer (pH
7) was used. (Diluent) The same diluent as above was used.

(Measurement method) The sample was diluted and introduced into the measurement system as in the following a), and measured under the measurement conditions in b). a) Dilution method of sample and introduction method to measurement system In Comparative Example 1, the measurement was performed using the apparatus shown in FIGS. 7 and 8. Using the sample suction nozzle 11, 1 μl of a measurement sample is collected, discharged into the dilution tank 13, and diluted with 50 μl of the diluent.
Diluted. Port the diluted sample to port d of the six-way valve 7
After being introduced into the sample loop 8 through the
Was connected as indicated by the broken line in FIG. 7, and 25 μl of the diluted sample was introduced into the separation column 3.

In the first embodiment, as described above, the measurement sample was diluted using the autosampler 6 in which the sample circuit was modified to have the structure shown in FIG. That is, as described with reference to FIG. 2D, 1 μl of a measurement sample was collected in the space 26 of the sample quantitative collection device 20 shown in FIG. 2D. Next, the measurement sample was diluted as described in the description of FIG. 4 and FIG. Specifically, the cleaning liquid 461 was introduced into the container 32, and the excess sample in the container 32 was washed away. The wastewater was discarded from the outlet 33. Next, as shown in FIG. 5A, the piston-like body 22 was pulled up until its lower end was located immediately above the sample suction port 25. Next, the measurement sample was diluted as described in the description of FIGS. 5 (b) to 5 (d). For this dilution, a total of 50 μl of diluent 471 was used. Pipeline 431-three-way valve 4 as shown in FIG.
3-Suction pump 10-The sample loop 8 is led through the three-way valve 44 to the six-way valve 7 (port d).
After that, the port of the six-way valve 7 was connected as shown by the broken line in FIG. 7, and 25 μl of the diluted sample was introduced into the separation column 3.

B) Measurement conditions The measurement conditions were eluent A (100 mM phosphate buffer, pH
Elution was performed with a step gradient using two liquids, 6) and eluent B (300 mM phosphate buffer, pH 7).

(Evaluation Method and Evaluation Results) From the obtained chromatogram, the total area of each peak was calculated by data processing. The same measurement sample was repeatedly measured 10 times, and the reproducibility of the total area was checked. Specifically, the average total area and the standard deviation were determined, and the coefficient of variation [(standard deviation / average total area) × 100] was determined from these. Table 1 shows the evaluation results obtained for each of Example 1 and Comparative Example 1.

[0034]

[Table 1]

Table 1 shows that the reproducibility of liquid chromatography was improved by diluting the measurement sample with an autosampler using the sample quantification and sampling apparatus of the present invention.

[0036]

The configuration of the sample quantification apparatus of the present invention is as described above. This sample quantification apparatus can accurately, simply and reproducibly collect even a very small amount of sample with high viscosity.
Therefore, for example, a more accurate chromatogram can be obtained by incorporating it into a sample dilution mechanism of a liquid chromatograph system.

[Brief description of the drawings]

FIG. 1 is a schematic explanatory view showing an example of a sample quantitative collection device of the present invention.

FIG. 2 is a diagram showing a method of using the sample quantitative collection device of FIG. 1;

FIG. 3 is an explanatory diagram in a case where the sample quantitative collection device is used continuously.

FIG. 4 is an explanatory diagram for describing an application example in which a sample is diluted in liquid chromatography using the sample quantitative collection device of FIG. 1;

FIG. 5 is a view for explaining a step of performing dilution using the apparatus having the configuration of FIG. 4;

FIG. 6 is an explanatory diagram for describing another application example of the sample quantitative collection device of the present invention.

FIG. 7 is a diagram showing a system configuration of a conventional liquid chromatograph.

FIG. 8 is a diagram showing a conventional sample suction nozzle.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Waste liquid container 2 Detector 3 Separation column 4 Liquid sending pump 5 Eluent 6 Autosampler 7 Hex valve 8 Sample loop 9 Waste liquid container 10 Suction pump 11 Sample suction nozzle 12 Sample 13 Dilution tank 20 Sample quantitative collection device 21 Cylindrical body 22 Piston-like body 23 Sample suction nozzle 24 Drive unit 25 Sample suction port 26 Space part 31 Projection 32 Container 33 Drain port 41 Three-way valve 42 Container 421 Cleaning liquid 43, 44, 45 Three-way valve 46 Container 461 Cleaning liquid 47 Container 471 Diluent 431 Piping a , B, c, d, e, f port

Claims (1)

[Claims] 1. A sample suction nozzle comprising a cylindrical body, a piston-like body accommodated therein and capable of reciprocating in a liquid-tight manner within the cylindrical body, and a projection having the same diameter as the piston-like body. An apparatus for collecting a fixed amount of a sample, comprising a container provided upright inside.