WO2013062084A1

WO2013062084A1 - Analysis device and analysis method

Info

Publication number: WO2013062084A1
Application number: PCT/JP2012/077717
Authority: WO
Inventors: 伊藤　伸也; 泉和氣
Original assignee: 株式会社日立ハイテクノロジーズ
Priority date: 2011-10-28
Filing date: 2012-10-26
Publication date: 2013-05-02
Also published as: JP2013096733A

Abstract

The present invention provides an analysis device and an analysis method with which samples containing drugs of different kinds and for various therapeutic ranges are subjected to a pretreatment under optimum conditions when the drug in the sample is measured. An analysis device (100) for measuring a drug contained in a measurement sample is provided with: an extract liquid injection mechanism (106) for injecting a solid-phase extract into a solid-phase extract cartridge (104); a pretreatment means for injecting a measurement sample into the solid-phase extract cartridge (104) into which the solid-phase extract has been injected, and generating an extract sample liquid by means of a solid extraction treatment; and a measurement means for measuring the drug in the extract sample liquid.

Description

Analysis apparatus and analysis method

The present invention relates to an analysis apparatus and an analysis method.

Conventionally, measurement of chemical substances present in minute amounts in liquid samples having a complicated composition such as biological samples such as blood, urine, and cell extracts has been performed. Such measurement methods include high-separation means such as liquid chromatography (Liquid Chromatography, LC) and capillary electrophoresis (Capillary Electrophoresis, CE), mass spectrometry (Mass Spectrometry, MS), and UV-visible absorption. Analytical methods combining detectors such as photometric methods (Ultraviolet-Visible-Spectrophotometry, UV) are used in many fields.

Mass spectrometry (MS) is an analysis method that ionizes a measurement target component and measures it based on the mass of the generated ions. An analysis method called MS / MS that performs mass analysis multiple times is a measurement target component ion. By fragmenting, a trace component in the sample solution can be detected with high sensitivity and high selectivity. LC-MS (or LC-MS / MS) in which liquid chromatography (LC) and mass spectrometry (MS) are connected on-line is widely used for analysis of trace substances in liquid samples.

In addition, pretreatments such as solid-phase extraction (Solid Phase Extraction, SPE) and liquid-liquid extraction (Liquid-Liquid Extraction, LLE) are used as means for increasing the degree of purification of the sample by LC-MS (or LC-MS / MS). It is done before the analysis. In particular, if a sample containing a major component with high viscosity and high concentration, such as blood, is injected directly into the LC-MS without pretreatment, not only will the measurement accuracy be reduced due to contamination of the device, but also due to clogging of the flow path. Measurement may not be possible. For this reason, pretreatment of a sample containing a main component having a high viscosity and a high concentration is essential.

Solid phase extraction (SPE) is present in a sample solution by capturing a chemical substance to be measured by interaction on a surface of a fine particle having a particle diameter of several μm to several tens of μm called a solid phase extraction material. It is a method of separating and purifying from other substances. In addition, liquid-liquid extraction (LLE) is a method in which a sample solution is dispensed and stirred in two types of solvents that are not mixed with each other, such as water and hexane, and the measurement target is determined by the difference in distribution coefficient between the two types of solvents. This is a method for separating a substance from other substances.
In particular, since solid-phase extraction (SPE) is easy to connect online with LC-MS (or LC-MS / MS), pretreatment by solid-phase extraction (SPE) and LC-MS (or LC-MS / MS) ) Can be automated. In addition, solid phase extraction (SPE) is characterized in that the pretreatment time can be reduced by simultaneous processing of multiple samples using a 96-well microplate or the like.

As a biological sample analysis using solid phase extraction (SPE) and LC-MS, for example, in Patent Document 1, biological samples such as serum and urine are processed off-line using a 96-well solid phase extraction plate and directly applied to MS. Quantification of amino acids, carnitines, sugars, immunosuppressants, and the like is performed by measuring by sample injection or LC-MS. In solid phase extraction (SPE), examples of the solid phase extraction cartridge used in offline processing include a 96-well plate described in Patent Document 1 and Patent Document 2, and a syringe container as described in Patent Document 3 and so on. Further, in the solid phase extraction (SPE), the solid phase extraction cartridge used by connecting online is the same as the precolumn of the liquid chromatography (LC) so as to withstand high pressure as described in Patent Document 4. There is something with the shape.

In any of the cartridges listed in Patent Document 1 to Patent Document 4, the solid phase extraction material is supplied in a state of being enclosed in the cartridge through a filter. As a means for forming a solid phase extraction cartridge at the time of measurement without being enclosed in a cartridge, Patent Document 5 provides a method of using a solid phase extraction material filled in a pipette tip provided with a filter inside.

US Patent Application Publication No. 2007/0004044 US Pat. No. 6,723,236 JP 2002-316002 A European Patent No. 11559597 US Pat. No. 6,770,246

In the automatic analyzer that measures the drug concentration in the blood, the type of drug to be measured varies depending on the sample, that is, the patient. Further, the concentration of the drug to be measured that is assumed to be present in the sample varies greatly depending on the type of drug. This is because the blood drug concentration (treatment area) where the therapeutic effect appears is different for each drug. As an example, in the case of sodium valproate used as an antiepileptic agent, the therapeutic range is 50 to 100 μg / mL, and the therapeutic range of tacrolimus used as an immunosuppressive agent is 2 to 20 ng / mL, which is 1000 times or more. There is a density difference.

That is, the automatic analyzer is required to measure various types of drugs in a wide range of concentrations assumed for each drug. For this purpose, it is required to perform an optimal pretreatment for each sample. Specifically, it is desired to optimize the amount of the sample solution, the type and amount of the extract, and the type and amount of the solid phase extraction material for each measurement target. Moreover, when an automatic analyzer is actually used for clinical examinations such as hospitals, when a highly urgent patient sample is supplied to the device, it is required to preferentially measure the sample.

In response to these requirements, the pretreatment apparatus using the 96-well plate described in Patent Document 1 and Patent Document 2 can process a large amount of samples at one time, while a dispensing head equipped with multiple syringes is provided. Since there is only one, there is a problem that a multi-item extraction process using different types or different amounts of eluate cannot be performed. Further, in clinical examinations, there is a problem that it is difficult to secure a sufficient amount of sample in a 96-well plate for each processing operation. Furthermore, in the case of a pre-processing apparatus using a plate, after starting processing once, the next processing request sample cannot be started until the processing being executed is completed, and it is difficult to perform flexible processing. There is a problem.

In the syringe-type solid phase extraction column described in Patent Document 3 and the cartridge-type column described in Patent Document 4, when changing the amount of the solid-phase extraction material, it is necessary to prepare as many columns in the apparatus as desired in advance. There is. For this reason, not only the cost of the consumables of the apparatus rises, but also a mechanism for selecting a plurality of types of columns for each sample is required, and there is a problem that the apparatus becomes complicated and large.

In the method described in Patent Document 5, the amount of the solid-phase extraction material can be changed for each sample by filling the pipette tip with the solid-phase extraction material slurry, but the solid-phase extraction material is prevented from dropping from the pipette tip. Therefore, it is necessary to use a viscous liquid such as glycerol as the slurry solvent. There is a problem in that this glycerol adsorbs on the surface of the extraction material, thereby affecting the solid-phase extraction efficiency of the drug. In particular, the drug to be measured is a chemical substance with various physicochemical properties depending on its use, and it is technically difficult to prepare a highly viscous liquid that does not affect all the drugs. . In addition, when a mass spectrometer is used as a detector, if a viscous liquid such as glycerol is introduced into the ion source, there is a problem in that the signal intensity decreases and the signal varies due to a decrease in ionization efficiency.

Therefore, the present invention provides an analyzer and an analysis method for performing pretreatment under optimal conditions for each sample for samples containing drugs of various types and therapeutic areas when measuring drugs in the sample. Is an issue.

In order to solve such a problem, the present invention is an analyzer for measuring a drug contained in a measurement sample, and is a solid phase extraction material dispensing means for injecting a solid phase extraction material into a solid phase extraction container Pretreatment means for injecting the measurement sample into the solid phase extraction container into which the solid phase extraction material has been injected and generating an extraction sample liquid by solid extraction processing, and the drug in the extraction sample liquid And a measuring means for measuring.

According to the present invention, when measuring a drug in a sample, pretreatment is performed on the sample containing various types and drugs in a therapeutic area under optimum conditions for each sample, and the measurement can be accurately performed in a mass spectrometer. An analysis apparatus and an analysis method that can be performed can be provided.

It is a block diagram of the automatic analyzer which concerns on 1st Embodiment. It is a chart which shows the process of the solid-phase extraction pretreatment by the automatic analyzer concerning a 1st embodiment. It is explanatory drawing which shows an example of the addition method of the sample information with respect to a sample container. It is sectional drawing which shows the structure of a solid-phase extraction cartridge. It is a block diagram of the automatic analyzer which concerns on 2nd Embodiment. It is a chart which shows the process of the solid phase extraction pretreatment by the automatic analyzer concerning a 2nd embodiment.

Hereinafter, modes for carrying out the present invention (hereinafter referred to as “embodiments”) will be described in detail with reference to the drawings as appropriate. In each figure, common portions are denoted by the same reference numerals, and redundant description is omitted.

<< First Embodiment >>
<Configuration of automatic analyzer 100>
FIG. 1 is a configuration diagram of an automatic analyzer 100 according to the first embodiment.
The automatic analyzer 100 is an analyzer that can automatically and continuously measure the concentration of a drug contained in a measurement sample. In the first embodiment, an asthma drug theophylline (drug in a serum sample (measurement sample) is used. ) Is analyzed for analysis.
The automatic analyzer 100 includes a sample container 101, a sample setting unit 102, a sample information reading unit 103, a solid phase extraction cartridge (solid phase extraction container) 104, an extraction material liquid container 105, and an extraction material liquid dispensing mechanism. 106, stirring mechanism 107, processing unit 108, reagent container 109, reagent placement unit 110, sample dispensing mechanism 111, reagent dispensing mechanism 112, pressurization mechanism 113, extraction container 114, and extraction The container installation unit 115, the consumable product installation unit 116, the sample introduction unit 117, the ion source 118, the mass analysis unit 119, and the control unit 120 are configured.

The sample container 101 stores a serum sample (measurement sample) to be measured. The sample placement unit 102 is configured such that the sample container 101 is disposed and the position of the sample vessel 101 can be moved (rotated) by rotating the sample placement unit 102. The sample information reading unit 103 can read the sample information written in the sample container 101. The solid phase extraction cartridge 104 dispenses a sample during the solid phase extraction process. The extraction material liquid container 105 stores the extraction material liquid. The extraction material liquid dispensing mechanism 106 is capable of dispensing the extraction material liquid slurry stored in the extraction material liquid container 105 to the solid phase extraction cartridge 104. The stirring mechanism 107 can form an extraction material liquid slurry by stirring the extraction material liquid in the extraction material liquid container 105. The processing unit 108 can sequentially perform a solid phase extraction process using the solid phase extraction cartridge 104. The reagent container 109 stores various reagents. The reagent placement unit 110 is provided with a reagent container 109 such as a washing solution or an eluate used for the extraction process, and the reagent placement unit 110 can be rotated to move the position of the reagent container 109 (rotation movement). It has become. The sample dispensing mechanism 111 is capable of dispensing a serum sample (measurement sample) stored in the sample container 101 to the solid phase extraction cartridge 104. The reagent dispensing mechanism 112 is capable of dispensing the reagent stored in the reagent container 109 to the solid phase extraction cartridge 104.

The pressurizing mechanism 113 can pass the liquid in the solid phase extraction cartridge 104 downward by pressurization. The extraction container 114 can collect the pretreated sample discharged from the solid phase extraction cartridge 104. The extraction container installation unit 115 is configured such that the extraction container 114 is arranged, and the position of the extraction container 114 can be moved (rotated) by the rotation of the extraction container installation unit 115. The consumables installation unit 116 is provided with consumables such as the solid phase extraction cartridge 104 and the extraction container 114. The sample introduction unit 117 can dispense the extracted sample and send the solution to the ion source 118. The ion source 118 is an electrospray ion source (ESI), and can ionize a sample that has been pretreated. The mass analyzer 119 can perform mass analysis on the components of the sample ionized by the ion source 118.
The control unit 120 stores sample information and measurement condition information, which will be described later, and can control the operation of the automatic analyzer 100 using these information. In addition, the control unit 120 can collect and analyze the results measured by the mass analysis unit 119.

Among the components of the automatic analyzer 100, some functions of the ion source 118, the mass analyzer 119, and the controller 120 are measurement means for measuring the concentration of the drug in the preprocessed measurement sample, and the remaining components (Including a part of the function of the control unit 120) is a pretreatment means for performing a pretreatment including a solid phase extraction process on the measurement sample.

<Concentration measurement by automatic analyzer 100>
Next, a procedure for measuring theophylline concentration in a serum sample by the automatic analyzer 100 will be described with reference to FIG. 1 and FIG. FIG. 2 is a chart showing steps of solid phase extraction pretreatment by the automatic analyzer 100 according to the first embodiment.
Prior to the processing of FIG. 2, a serum sample (measurement sample) is dispensed and stored in the sample container 101 and set in the sample setting unit 102. At this time, for example, a bar code 121 in which information (sample information) related to the sample such as the specimen ID, the liquid property of the sample, and the drug to be measured is recorded is prepared in advance. A code 121 is attached.

Returning to the description of FIG. 2, the automatic analyzer 100 measures the theophylline concentration in the serum sample in the following steps.
(Step 1: Read sample information)
The automatic analyzer 100 rotates the sample setting unit 102 in which the sample container 101 is set, and passes the sample container 101 in front of the sample information reading unit 103. The sample information reading unit 103 reads sample information from the barcode 121 of the sample container 101 and sends the read information to the control unit 120.

(Process 2: Call preconditions)
The control unit 120 includes, for each sample type and drug, pretreatment conditions such as sample amount, type and amount of extraction material, solid-phase extraction conditions (such as type and amount of washing liquid and eluate, pressurization condition), and mass. Analysis (MS) measurement conditions are stored in advance as a database DB. When there are a plurality of sample types used for concentration analysis for a certain drug, a database is created for each sample type for that drug. The control unit 120 calls the preprocessing conditions and mass spectrometry (MS) measurement conditions used for theophylline measurement from the sample information sent from the sample information reading unit 103 as device control parameters, and transmits them to each unit of the device.

In the case of theophylline, the therapeutic range is 10 to 20 μg / mL, and the automatic analyzer 100 needs to quantify in a concentration range of about 1 to 50 μg / mL including the therapeutic range. This concentration range is a considerably high concentration range in view of detection sensitivity of mass spectrometry (MS) for theophylline, and therefore, a small amount of sample may be used, for example, about 20 μL is sufficient. Similarly, parameters relating to the amount of the solid-phase extraction material optimized for theophylline measurement in the assumed concentration range, the washing solution, and the eluate are called and transmitted to each part of the apparatus.

(Process 3: Extraction material liquid slurry injection)
The extraction material liquid dispensing mechanism 106 dispenses a set amount of the extracted material liquid slurry from the extraction material liquid container 105 based on the device control parameter transmitted from the control unit 120, and supplies it to the solid phase extraction cartridge 104. inject. The extraction material liquid is an organic solvent such as water or methanol containing a solid phase extraction material. As the solid phase extraction material, for example, “NOBIAS RP-SG1” manufactured by Hitachi High-Technologies Corporation, which is a reverse phase solid phase extraction material, can be used. In this case, a 50% aqueous methanol solution is suitable as the solvent. The ratio of the extraction material liquid slurry is suitably 1 mL of 50% methanol aqueous solution with respect to 20 mg of the solid phase extraction material.

The extraction material liquid slurry is sufficiently stirred before dispensing by the stirring mechanism 107 to form a uniform slurry of the solid phase extraction material. As the stirring mechanism 107, a magnetic stirrer is simple. In the magnetic stirrer, a stirrer covered with a Teflon (registered trademark) resin or the like with a bar magnet is put into a substance to be stirred (in this case, an extractant liquid slurry), and installed in the lower part of the extractant liquid container 105. The magnet inside the stirring mechanism 107 is rotated by a motor. By stirring with a magnetic stirrer, a uniform slurry can be formed. The solid phase extraction cartridge 104 into which the extraction material liquid slurry has been injected is pressurized from above by the pressurizing mechanism 113, and the solvent passes downward.

FIG. 4 is a cross-sectional view showing the structure of the solid phase extraction cartridge 104. A filter 122 is installed below the inside of the solid-phase extraction cartridge 104. In the present embodiment, the hole diameter of the filter 122 is 10 μm. According to such a structure, the solvent pressurized by the pressurizing mechanism 113 passes through the solid phase extraction cartridge 104, while the solid phase extraction material remains on the filter 122.

(Step 4: Equilibration)
In order to equilibrate the solid phase extraction material, pure water arranged in the reagent arrangement unit 110 is dispensed by the reagent dispensing mechanism 112 to the solid phase extraction cartridge 104 after passing through the solvent. The injected pure water is pressurized by the pressurizing mechanism 113 and passes below the solid phase extraction cartridge 104. The preparation for sample processing is completed by the steps so far.

(Step 5: Sample injection)
The reagent dispensing mechanism 112 dispenses a theophylline internal standard substance (theophylline-d6) solution from the reagent placement unit 110 and injects it into the solid phase extraction cartridge 104 in which the solid phase extraction material has been injected and the sample processing preparation has been completed. Subsequently, the sample dispensing mechanism 111 dispenses a serum sample (measurement sample) from the sample container 101 and injects it into the solid phase extraction cartridge 104 into which the internal standard substance has been injected. The serum sample and the internal standard substance liquid injected into the solid phase extraction cartridge 104 are pressurized from above by the pressurizing mechanism 113, pass through the solid phase extraction material, and discharged downward. By this step, theophylline (drug) and the internal standard substance contained in the serum sample are selectively adsorbed on the surface of the solid-phase extraction material by hydrophobic interaction.

(Step 6: Washing)
The reagent dispensing mechanism 112 dispenses 200 μL of pure water as a cleaning liquid stored in the reagent container 109. Then, the solid-phase extraction cartridge 104 is washed by performing a pressure treatment by the pressure mechanism 113. By this step, components in the serum sample remaining nonspecifically on the surface of the solid-phase extraction material are discharged.

(Step 7: Elution)
After washing, the reagent dispensing mechanism 112 dispenses 100 μL of 50% methanol aqueous solution as an eluent into the solid phase extraction cartridge 104. The extraction sample solution in which theophylline (drug) and theophylline-d6 (internal standard substance) are eluted from the solid-phase extraction cartridge 104 is collected in the extraction container 114 by performing a pressurizing process with the pressurizing mechanism 113.

The automatic analyzer 100 moves the extraction container 114 from which the extraction sample solution has been collected to the position of the sample introduction unit 117 by the rotation of the extraction container installation unit 115. The extracted sample solution is introduced into the ion source 118 by the sample introduction unit 117. Theophylline (drug) and theophylline-d6 (internal standard substance) are ionized by the ion source 118, and both components are detected by the mass spectrometer 119. The detected signal strengths of both components are transferred to the control unit 120. The control unit 120 calculates the theophylline concentration in the serum sample from the transferred signal intensity ratio of both components and the signal intensity ratio of the calibrator obtained in advance.
In the process as described above, the automatic analyzer 100 measures the theophylline concentration in the serum sample.

<Summary>
As described above, when the automatic analyzer 100 performs quantitative measurement of various types of drugs, the pretreatment conditions corresponding to the types of each drug and the sample to be measured are stored in a database, and according to the values of the database. Pretreatment including solid phase extraction is performed. As a result, it is possible to always select the optimal amount of solid phase extraction material, sample, and various reagents, to suppress unnecessary reagent consumption and to reduce the running cost for analysis processing.

In addition, according to the analyzer of the prior art, it has been necessary to provide a plurality of different solid phase extraction cartridges for each type and amount of solid phase extraction material. On the other hand, in the automatic analyzer 100, since it is not necessary to provide a solid phase extraction cartridge in which a solid phase extraction material is sealed in advance, the apparatus configuration can be simplified and the manufacturing cost of the apparatus itself can be reduced.

<< Second Embodiment >>
<Configuration of automatic analyzer 500>
FIG. 5 is a configuration diagram of an automatic analyzer 500 according to the second embodiment.
The automatic analyzer 500 according to the second embodiment is different from the automatic analyzer 100 according to the first embodiment in that a stirring mechanism 123 that stirs the inside of the container with respect to the solid phase extraction cartridge 104 is provided. Yes.
As the stirring mechanism 123, for example, an ultrasonic stirrer can be used. By providing the stirring mechanism 123, the contact rate between the sample in the solid-phase extraction cartridge 104 and a reagent such as a solid-phase extraction material can be improved. For example, the concentration of the measurement target substance (drug) in the measurement sample can be increased. Even if it is low, it is possible to measure accurately.

The automatic analyzer 500 is an analyzer that can automatically and continuously measure the concentration of a drug contained in a measurement sample. In the second embodiment, the immunosuppressant tacrolimus (drug) contained in a whole blood sample is used. It is assumed that the concentration is analyzed.
Since tacrolimus in the whole blood sample is present in blood cell components, solid phase extraction cannot be performed as it is. In the second embodiment, a methanol solution containing 3 times the amount of 0.5 M zinc sulfate aqueous solution and 4 times the amount of 10 ng / mL concentration of ascomachine (0.5 M sulfuric acid for 50 μL of the whole blood sample). By adding 150 μL of aqueous zinc solution and 200 μL of methanol solution, blood cells are destroyed and precipitates are formed, and the supernatant after centrifugation is used as a whole blood treatment liquid sample (measurement sample). Here, ascomycin is used at the time of mass spectrometry (MS) measurement as an internal standard substance of tacrolimus.

Further, in the second embodiment, the solid phase extraction process is performed in a batch manner in the pretreatment. Here, the batch type refers to a system in which solid phase extraction is performed with the drug in the measurement sample by freely moving the measurement sample solution without fixing the solid phase extraction material to a part of the cartridge.

<Concentration measurement by automatic analyzer 500>
Next, a procedure for measuring tacrolimus concentration in a whole blood sample by the automatic analyzer 500 will be described with reference to FIG. 5 and FIG. FIG. 6 is a chart showing steps of solid phase extraction pretreatment by the automatic analyzer 500 according to the second embodiment.
Prior to the processing of FIG. 6, a whole blood processing solution sample (measurement sample) is dispensed and stored in the sample container 101, set in the sample setting unit 102, and a barcode 121 in which sample information is recorded (see FIG. 3) ) Is attached to the side surface of the sample container 101.

(Step 1: Read sample information)
This is performed in the same manner as (Step 1: Reading sample information) (see FIG. 2) of the first embodiment.

(Process 2: Call preconditions)
The control unit 120 calls the preprocessing conditions and mass spectrometry (MS) measurement conditions used for tacrolimus measurement as device control parameters from the sample information sent from the sample information reading unit 103, and transmits them to each unit of the device.

In the case of tacrolimus, the therapeutic range is 2 to 20 ng / mL, and the automatic analyzer 500 is required to perform quantification in a concentration range of about 0.5 to 30 ng / mL including the therapeutic range. Thus, tacrolimus in whole blood has a lower concentration than, for example, theophylline mentioned in the first embodiment, and a sample amount of 200 μL is required for accurate measurement by the automatic analyzer 500. Similarly, parameters relating to the amount of the solid-phase extraction material optimized for the measurement of tacrolimus in the assumed concentration range, the washing liquid, the eluate, etc. are called and transmitted to each part of the apparatus.

(Process 3: Extraction material liquid slurry injection)
Similar to the (step 3: extraction material liquid slurry injection) of the first embodiment (see FIG. 2), the extraction material liquid dispensing mechanism 106 is set based on the apparatus control parameter transmitted from the control unit 120. A certain amount of extraction material liquid slurry is dispensed from the extraction material liquid container 105 and injected into the solid phase extraction cartridge 104. The solid-phase extraction cartridge 104 into which the extraction material liquid slurry has been injected is pressurized from above by the pressurizing mechanism 113, and the solvent passes downward.

(Step 5A: Sample injection)
The sample dispensing mechanism 111 dispenses the whole blood processing liquid sample (measurement sample) from the sample container 101 and injects it into the solid phase extraction cartridge 104 into which the solid phase extraction material has been injected.

(Step 5B: stirring)
The automatic analyzer 100 rotates the processing unit 108 to move the solid-phase extraction cartridge 104 into which the solid-phase extraction material and the whole blood processing liquid sample (measurement sample) are dispensed to the position of the stirring mechanism 123, thereby Stir the phase extraction material and the whole blood treatment liquid sample. By this stirring, the solid phase extraction material, tacrolimus in the sample solution and the internal standard ascomycin can be brought into contact with each other a sufficient number of times, and are adsorbed on the solid phase extraction material.

(Process 5C: Discharge)
The automatic analyzer 100 rotates the processing unit 108 to move the solid-phase extraction cartridge 104 that has been sufficiently stirred to the position of the pressurizing mechanism 113. The whole blood processing liquid sample in the solid-phase extraction cartridge 104 passes downward by the pressurizing process of the pressurizing mechanism 113. On the other hand, tacrolimus and ascomycin remain adsorbed on the solid phase extraction material, and remain on the upper part of the filter 122 in the cartridge together with the solid phase extraction material.

(Step 6: Washing)
Similar to (Step 6: Washing) of the first embodiment (see FIG. 2), the reagent dispensing mechanism 112 dispenses 200 μL of pure water as a washing liquid stored in the reagent container 109. Then, the solid-phase extraction cartridge 104 is washed by performing a pressure treatment by the pressure mechanism 113. By this step, the components in the whole blood processing liquid sample remaining non-specifically on the surface of the solid-phase extraction material are discharged.

(Step 7: Elution)
After washing, the reagent dispensing mechanism 112 dispenses 50 μL of methanol as an eluent to the solid phase extraction cartridge 104. Then, the extraction sample solution in which tacrolimus (drug) and ascomycin (internal standard substance) are eluted from the solid-phase extraction cartridge 104 is recovered in the extraction container 114 by performing a pressurizing process by the pressurizing mechanism 113.

The automatic analyzer 500 moves the extraction container 114 from which the extraction sample solution has been collected to the position of the sample introduction unit 117 by the rotation of the extraction container installation unit 115. The extracted sample solution is introduced into the ion source 118 by the sample introduction unit 117. Tacrolimus (drug) and ascomycin (internal standard substance) are ionized by the ion source 118, and both components are detected by the mass spectrometer 119. The detected signal strengths of both components are transferred to the control unit 120. The control unit 120 calculates the tacrolimus concentration in the whole blood sample from the transferred signal intensity ratio of both components and the signal intensity ratio of the calibrator obtained in advance.
In the process as described above, the automatic analyzer 500 measures the tacrolimus concentration in the whole blood sample.

As described above, according to the automatic analyzer 500, in addition to the same effects as those of the automatic analyzer 100 according to the first embodiment, it is possible to accurately measure even when the concentration of the measurement target substance in the sample is low. it can.

≪Modification≫
The automatic analyzer according to the present embodiment is not limited to the configuration of the above-described embodiment, and various modifications can be made without departing from the spirit of the invention.
In the first and second embodiments, the measurement using a single solid phase extraction material has been described, but it is also possible to supply a plurality of solid phase extraction materials to one solid phase extraction cartridge. Specifically, a plurality of extraction material liquid containers 105 each containing different types of solid phase extraction materials are arranged on the operation line of the extraction material liquid dispensing mechanism 106, and a plurality of (Step 3: extraction material liquid slurry injection) are arranged. It can be realized by repeating it. Such a method is effective, for example, when measuring a drug whose solid-phase extraction efficiency is improved by mixing a cation exchange solid-phase extraction material having a sulfonic acid group and a reverse-phase solid-phase extraction material. . In the case of adopting such a method, it is necessary to optimize the pH of the washing solution or the eluate and to change the type and amount of the eluate, but these changes are easy if the

automatic analyzers

100 and 500 are used.

Although the mass spectrometer 119 of the

automatic analyzers

100 and 500 according to the present embodiment has been described as performing quantitative analysis for measuring the concentration of a drug in the extracted sample solution, the present invention is not limited to this, and the extracted sample solution is not limited thereto. A qualitative analysis for identifying the substance of the drug in the drug may be performed, or a qualitative analysis and a quantitative analysis may be performed simultaneously.

The analyzer according to the present invention is effective as an analyzer for measuring the concentration of a drug contained in a measurement sample, and particularly using a mass spectrometer such as an LC-MS (or LC-MS / MS) analyzer. It is suitable for an analyzer that measures the concentration of a drug contained in a biological sample.

100 Automatic analyzer (analyzer)
101 Sample container 102 Sample installation part (pretreatment means)
103 Sample information reading unit (pretreatment means)
104 Solid phase extraction cartridge (solid phase extraction container)
105 Extraction material liquid container (slurry generating means)
106 Extraction material liquid dispensing mechanism (Solid phase extraction material dispensing means)
107 Stirring mechanism (slurry generating means)
108 processing unit (pre-processing means)
109 Reagent container 110 Reagent placement part (pretreatment means)
111 Sample dispensing mechanism (pretreatment means)
112 Reagent dispensing mechanism (pretreatment means)
113 Pressurizing mechanism (pretreatment means)
114 Extraction container 115 Extraction container installation part (pretreatment means)
116 Consumables installation unit 117 Sample introduction unit (pretreatment means)
118 Ion source (measuring means)
119 Mass Spectrometer (Measuring means)
120 control part 123 stirring mechanism (stirring means)
500 Automatic analyzer (analyzer)

Claims

An analyzer for measuring a drug contained in a measurement sample,
A solid phase extraction material dispensing means for injecting the solid phase extraction material into the solid phase extraction container;
Pretreatment means for injecting the measurement sample into the solid phase extraction container into which the solid phase extraction material has been injected and generating an extraction sample liquid by solid extraction processing;
And an analyzer that measures the drug in the extracted sample solution.
A slurry generation means for stirring the solid phase extraction material and the solvent to generate a solid phase extraction material slurry,
The solid phase extraction material dispensing means includes:
The analyzer according to claim 1, wherein the solid phase extraction material slurry generated by the slurry generating means is injected into the solid phase extraction container.
The preprocessing means includes
2. The analyzer according to claim 1, further comprising a stirring unit that stirs the solid-phase extraction material and the measurement sample injected into the solid-phase extraction container in the solid-phase extraction container.
The analyzer according to claim 1, wherein the measuring unit performs a qualitative analysis for identifying a substance of the drug in the extracted sample solution.
The analyzer according to claim 1, wherein the measuring unit performs a quantitative analysis for measuring the concentration of the drug in the extracted sample solution.
The solid phase extraction material dispensing means includes:
6. The analyzer according to claim 5, wherein an amount of the solid phase extraction material injected into the solid phase extraction container is changed based on an assumed concentration of the drug contained in the measurement sample. .
The preprocessing means includes
The analyzer according to claim 5, wherein the amount of the measurement sample injected into the solid phase extraction container is changed based on an assumed concentration of the drug contained in the measurement sample.
The solid phase extraction material dispensing means includes:
The analyzer according to claim 1, wherein a plurality of types of solid phase extraction materials are injected into one said solid phase extraction container.
The analyzer according to claim 1, wherein the measuring unit is a mass spectrometer.
The analyzer according to claim 1, wherein the measurement sample is a biological fluid including blood, serum, urine, and saliva.
There is an analysis method by an analyzer for measuring a drug contained in a measurement sample,
An injection process for injecting a solid-phase extraction material into a solid-phase extraction container;
A pre-process for injecting the measurement sample into the solid-phase extraction container into which the solid-phase extraction material has been injected by the injection process and generating an extraction sample liquid by a solid extraction process;
And performing a measurement process for measuring the drug in the extracted sample solution by the pretreatment.