JP5941092B2 - Analysis apparatus and analysis method - Google Patents

Description

  The present invention relates to an analysis apparatus and an analysis method using, for example, capillary electrophoresis.

  An analysis method for analyzing the concentration or amount of a specific component contained in a sample includes a method having a separation step for separating the specific component from the sample and a detection step for detecting the separated specific component. For example, in an analysis method using capillary electrophoresis, a separation channel having a relatively small cross-sectional area is filled with an electrophoresis solution, and the sample is introduced near one end of the separation channel. When voltage is applied to both ends of the separation channel, an electroosmotic flow is generated in which the electrophoresis solution moves from the positive electrode side to the negative electrode side by electrophoresis. In addition, when the voltage is applied, the specific component tends to move according to the electrophoretic mobility. Accordingly, the specific component moves according to a velocity vector obtained by synthesizing the velocity vector of the electroosmotic flow and the velocity vector of movement by electrophoresis. By this movement, the specific component is separated from other components. By detecting the separated specific component by, for example, an optical method, the amount and concentration of the specific component can be analyzed.

  When the analysis method is performed on a plurality of samples, a plurality of the separation channels are prepared, and these separation channels are repeatedly used. If the next analysis is performed while the sample remains in the separation channel after the previous analysis, a correct analysis result cannot be obtained. For this reason, it is necessary to wash the separation channel every time analysis is completed. FIG. 8 shows a timing chart in the conventional analysis method (see, for example, Patent Document 1). In this analysis method, four separation channels 91a, 91b, 91c, and 91d are used.

  First, a pretreatment step 92a including a cleaning step is performed on the separation channel 91a. In the separation channel 91a, an analysis step 93a using capillary electrophoresis is performed following the pretreatment step 92a. In addition, after the pretreatment step 92a is completed, the pretreatment step 92b and the analysis step 93b are subsequently performed in the separation channel 91b. In addition, after the pretreatment step 92b is completed, the pretreatment step 92c and the analysis step 93c are subsequently performed in the separation channel 91c. Then, after the pretreatment step 92c is completed, the pretreatment step 92d and the analysis step 93d are subsequently performed in the separation channel 91d. Thus, in this analysis method, the pretreatment steps 92a, 92b, 92c, and 92d are continuously performed as a series of steps. When analyzing a large number of samples, the process shown in this figure is repeated. Thereby, for example, it is not necessary to wait for the start of the pretreatment process 92b until the analysis process 93a is completed, and the time required to complete all the analysis processes is reduced.

  However, when more samples are targeted, it is required to perform the analysis more efficiently. In the above analysis method, for example, in the separation channel 91b, the analysis step 93b cannot be started unless a time obtained by adding the time for completing the pretreatment step 92a and the time for completing the pretreatment step 92b has elapsed. Thus, in the above analysis method, the waiting time until the analysis steps 93a, 93b, 93c, and 93d are started hinders shortening of the entire analysis time.

Japanese Patent No. 4375031

  The present invention has been conceived under the circumstances described above, and it is an object of the present invention to provide an analysis apparatus and an analysis method capable of analyzing more efficiently.

  The analyzer provided by the first aspect of the present invention includes a plurality of separation channels for separating a specific component contained in an introduced sample, detection means for detecting the separated specific component, and each of the separations. A control unit that controls a separation step that performs the separation in the flow channel and an analysis step that includes the detection step that performs the detection, and a pretreatment step that sets the separation flow channels in a state where the analysis step can be performed The control unit simultaneously executes at least a part of each of the pretreatment steps of at least two separation channels among the plurality of separation channels.

  In a preferred embodiment of the present invention, the pretreatment step includes a washing step for washing the separation channels.

  In a preferred embodiment of the present invention, a cleaning liquid tank storing a cleaning liquid used in the cleaning process is further provided, and the plurality of separation flow paths are connected to the cleaning liquid tank via branch flow paths. The control unit controls which of the plurality of separation flow paths the cleaning liquid flows from the cleaning liquid tank.

  In a preferred embodiment of the present invention, the branch flow path is provided with a switching valve for switching which of the plurality of separation flow paths the cleaning liquid flows from the cleaning liquid tank. The operation is controlled by the control unit.

  In a preferred embodiment of the present invention, the separation step utilizes electrophoresis, and an electrophoresis solution tank storing an electrophoresis solution used for electrophoresis is further connected to the upstream side of the switching valve. The control unit selectively introduces the washing solution or the electrophoresis solution into any of the plurality of separation channels by the switching operation of the switching valve.

  In a preferred embodiment of the present invention, the pretreatment step further includes a filling step of filling the separation channel after the washing step with the electrophoresis solution.

  In a preferred embodiment of the present invention, the control unit sets the start time of the pretreatment process or the start time of the analysis process of the plurality of separation channels to different times.

  In a preferred embodiment of the present invention, the detection means comprises a plurality of detection units each provided in each of the separation channels.

  In a preferred embodiment of the present invention, each of the detection units performs detection at a position biased to either end from the center of each of the separation channels, and each of the separation channels has any of its both ends. The sample can also be introduced.

  In a preferred embodiment of the present invention, pressure generating means for applying a pressure capable of discharging the liquid filled in each of the separation channels is connected to the plurality of separation channels.

  In a preferred embodiment of the present invention, a manifold for equalizing pressure applied to the plurality of separation channels is provided between the plurality of separation channels and the pressure generating means. .

  In a preferred embodiment of the present invention, it further comprises a dispensing means having a nozzle for dispensing the sample into each separation channel, and the pretreatment step dispenses the sample into the separation channel. It further includes a dispensing step.

  In a preferred embodiment of the present invention, the apparatus further includes a sample container having a lid for containing the sample and blocking the sample from the outside air, and the nozzle can penetrate the lid.

  In a preferred embodiment of the present invention, the pretreatment step includes a dilution step in which the sample is diluted with a diluent in a dilution tank, and the sample and the sample are sucked into and discharged from the nozzle. Stir the diluted solution.

  In a preferred embodiment of the present invention, the separation step utilizes electrophoresis, and the electrophoresis solution used for electrophoresis also serves as the diluent.

  In a preferred embodiment of the present invention, each of the separation channels has a pair of electrodes provided near both ends, and a common voltage is applied to cause the electrophoresis in each of the separation channels. And a switch capable of selecting which one of the pair of electrodes of the plurality of separation flow paths is electrically connected, and the switching operation of the switch is performed by the control unit. Be controlled.

  In a preferred embodiment of the present invention, the power supply unit can switch the polarity of the voltage applied to each separation channel.

  In a preferred embodiment of the present invention, the cross section of each separation channel is a circle having a diameter of 25 to 100 μm or a rectangle having a side length of 25 to 100 μm.

  In a preferred embodiment of the present invention, the sample contains hemoglobin.

  In a preferred embodiment of the present invention, the sample is blood.

  In a preferred embodiment of the present invention, the control unit averages the analysis results in the separation channels when the analysis step is performed in two or more separation channels for the same sample. To do.

  In preferable embodiment of this invention, the said control part performs the said averaging process except for what was judged to be abnormal among the analysis results of the said 2 or more separation flow paths.

  In a preferred embodiment of the present invention, the control unit performs a correction calculation process on the analysis result using a correction coefficient set in each separation channel.

  The analysis method provided by the second aspect of the present invention uses a plurality of separation channels to separate a specific component of a sample introduced into each of the separation channels, and the separated specific component An analysis step including a detection step detected by a detection means, and a pretreatment step for making each of the separation channels into a state where the analysis step can be performed, and at least two of the plurality of separation channels At least a part of each of the pretreatment steps of the two separation channels is performed simultaneously.

  In a preferred embodiment of the present invention, the pretreatment step includes a washing step for washing the separation channels.

  In a preferred embodiment of the present invention, the plurality of separation channels are connected via a branch channel to a cleaning liquid tank in which a cleaning liquid used in the cleaning step is stored. The cleaning liquid is selectively allowed to flow from the cleaning liquid tank toward either side.

  In a preferred embodiment of the present invention, the branch flow path is provided with a switching valve for switching which of the plurality of separation flow paths the cleaning liquid flows from the cleaning liquid tank.

  In a preferred embodiment of the present invention, the separation step utilizes electrophoresis, and an electrophoresis solution tank storing an electrophoresis solution used for electrophoresis is further connected to the upstream side of the switching valve. The washing solution or the electrophoresis solution is selectively introduced into any of the plurality of separation channels by the switching operation of the switching valve.

  In a preferred embodiment of the present invention, the pretreatment step further includes a filling step of filling the separation channel after the washing step with the electrophoresis solution.

  In a preferred embodiment of the present invention, the start time of the pretreatment step or the start time of the analysis step of the plurality of separation channels is set to different times.

  In a preferred embodiment of the present invention, the detection means comprises a plurality of detection units each provided in each of the separation channels.

  In a preferred embodiment of the present invention, each of the detection units performs detection at a position biased to either end from the center of each of the separation channels, and each of the separation channels has any of its both ends. The sample can also be introduced.

  In a preferred embodiment of the present invention, pressure generating means for applying a pressure capable of discharging the liquid filled in each of the separation channels is connected to the plurality of separation channels.

  In a preferred embodiment of the present invention, a manifold for equalizing pressure applied to the plurality of separation channels is provided between the plurality of separation channels and the pressure generating means. .

  In a preferred embodiment of the present invention, the pretreatment step further includes a dispensing step of dispensing the sample into the separation channel using a dispensing means having a nozzle.

  In a preferred embodiment of the present invention, the sample is sucked into the syringe by allowing the nozzle to pass through a lid provided in a sample container for containing the sample, for blocking the sample from outside air. .

  In a preferred embodiment of the present invention, the pretreatment step includes a dilution step in which the sample is diluted with a diluent in a dilution tank, and the sample and the sample are sucked into and discharged from the nozzle. Stir the diluted solution.

In a preferred embodiment of the present invention, the separation step utilizes electrophoresis,
The electrophoresis solution used for electrophoresis also serves as the diluent.

  In a preferred embodiment of the present invention, each of the separation channels has a pair of electrodes provided near both ends, and the switch of the common power source unit is used to switch the switch. A voltage capable of causing electrophoresis is selectively applied to the pair of electrodes.

  In a preferred embodiment of the present invention, the power supply unit can switch the polarity of the voltage applied to each separation channel.

  In a preferred embodiment of the present invention, the cross section of each separation channel is a circle having a diameter of 25 to 100 μm or a rectangle having a side length of 25 to 100 μm.

  In a preferred embodiment of the present invention, the sample contains hemoglobin.

  In a preferred embodiment of the present invention, the sample is blood.

  In a preferred embodiment of the present invention, when the analysis step is performed in two or more separation channels for the same sample, the analysis results in these separation channels are averaged.

  In a preferred embodiment of the present invention, the averaging process is performed except for the analysis results of the two or more separation channels that are determined to be abnormal.

  In a preferred embodiment of the present invention, correction calculation processing is performed on the analysis result using the correction coefficient set for each separation channel.

  According to such a configuration, the time required to complete the pretreatment process of the plurality of separation channels is clearly shorter than the total time of completing the pretreatment processes individually. For this reason, it is possible to finish the said analysis process quite early compared with the example shown, for example in FIG. Thereby, the analysis using the said analyzer can be performed more efficiently.

  Other features and advantages of the present invention will become more apparent from the detailed description given below with reference to the accompanying drawings.

It is a system schematic diagram showing an example of an analysis device concerning the present invention. It is a circuit diagram which shows the power supply structure of the analyzer of this embodiment. It is a flowchart which shows an example of the analysis method which concerns on this invention. It is the schematic which shows the process of extracting a sample from a sample container. It is the schematic which shows the dilution and stirring process in a dilution tank. It is the schematic which shows the fractionation process to a separation channel. It is a timing chart which shows an example of the analysis method concerning the present invention. It is a timing chart which shows an example of the conventional analysis method.

  Hereinafter, preferred embodiments of the present invention will be specifically described with reference to the drawings.

  FIG. 1 shows an example of an analyzer according to the present invention. The analyzer A of the present embodiment includes a storage tank unit 10, a dispensing unit 20, a plurality of separation channels 30a, 30b, 30c, and 30d, a detection unit 40, a control unit 71, and a power supply unit 72. In the present embodiment, the analyzer A performs analysis using capillary electrophoresis. In FIG. 1, the power supply unit 72 is omitted for the sake of understanding.

  The storage tank unit 10 includes an electrophoretic liquid tank 11, a purified water tank 12, and a cleaning liquid tank 13. In the electrophoretic solution tank 11, an electrophoretic solution L1 is stored. The electrophoresis liquid L1 is a liquid that functions as a so-called buffer in capillary electrophoresis, and is, for example, a 100 mM malic acid-arginine buffer (pH 5.0). The purified water tank 12 stores purified water L2. The cleaning liquid L3 is stored in the cleaning liquid tank 13.

  The dispensing means 20 has a function of dispensing the sample Sp into a plurality of separation channels 30a, 30b, 30c, and 30d, and further diluting the sample Sp into a state suitable for analysis. The dispensing means 20 includes a sample container 23, a dilution tank 25, a syringe 21, and a nozzle 22.

  The sample container 23 is a glass blood collection tube, for example, and accommodates a sample Sp such as whole blood. For example, a rubber lid 24 is fitted in the sample container 23. The dilution tank 25 is a place for diluting the sample Sp, which is whole blood, to a concentration suitable for analysis. The syringe 21 can be inhaled and ejected, and is connected to a nozzle 22. The nozzle 22 is a part through which the sample Sp is sucked and discharged by the suction and discharge movements of the syringe 21. In the present embodiment, the nozzle 22 is made of stainless steel, for example, and has a sharp shape with its tip cut obliquely. The nozzle 22 is supported by a drive mechanism (not shown). By this drive mechanism, the nozzle 22 is inserted into and extracted from the sample container 23, entered and exited from the dilution tank 25, and the inlets 31a, 31b, 31c, 31d of the separation channels 30a, 30b, 30c, 30d. It is possible to enter and exit. Further, the nozzle 22 may be capable of entering and exiting the discharge holes 32a, 32b, 32c, and 32d.

  The plurality of separation channels 30a, 30b, 30c, and 30d are places where analysis using capillary electrophoresis is performed, and are fine channels formed in the main bodies 36a, 36b, 36c, and 36d made of silica, for example. is there. The cross-sections of the separation channels 30a, 30b, 30c, and 30d are preferably circular with a diameter of 25 to 100 μm or rectangular with a side length of 25 to 100 μm, but are suitable for performing capillary electrophoresis. The shape and dimensions are not limited to this. In the present embodiment, the length of the separation channels 30a, 30b, 30c, and 30d is about 30 mm, but is not limited to this.

  In the separation flow paths 30a, 30b, 30c, 30d, introduction holes 31a, 31b, 31c, 31d and discharge holes 32a, 32b, 32c, 32d are formed. The introduction holes 31a, 31b, 31c, and 31d are provided at one end of the separation channels 30a, 30b, 30c, and 30d, and are portions into which the sample Sp is introduced. In the present embodiment, the electrophoresis solution L1, the purified water L2, and the cleaning solution L3 can be introduced. The discharge holes 32a, 32b, 32c, and 32d are provided at the other ends of the separation channels 30a, 30b, 30c, and 30d, and the sample Sp, electrophoresis solution L1, purified water L2, and This is a portion where the cleaning liquid L3 and the like are discharged.

  The separation channels 30a, 30b, 30c, and 30d are provided with electrodes 33a, 33b, 33c, and 33d and electrodes 34a, 34b, 34c, and 34d. In the present embodiment, the electrodes 33a, 33b, 33c and 33d are exposed to the introduction holes 31a, 31b, 31c and 31d, and the electrodes 34a, 34b, 34c and 34d are the discharge holes 32a, 32b, 32c and 32d. Is exposed.

  The detection unit 40 is for analyzing a specific component separated from other components of the sample Sp in the separation channels 30a, 30b, 30c, and 30d, and includes detection units 41a, 41b, 41c, and 41d. The detection units 41a, 41b, 41c, and 41d are located in portions of the separation channels 30a, 30b, 30c, and 30d that are closer to the discharge holes 32a, 32b, 32c, and 32d than the introduction holes 31a, 31b, 31c, and 31d. Is provided. The detectors 41a, 41b, 41c, 41d are each composed of, for example, a light source (not shown) and a light receiving part (not shown). The sample Sp is irradiated with light from the light source, and reflected light from the sample Sp is received by the light receiving unit. Thereby, the light absorbency of sample Sp is measured.

  The control unit 71 is for controlling the operation of each unit of the analyzer A, and performs a series of controls for realizing analysis by the analyzer A. The control unit 71 includes, for example, a CPU, a memory, an input / output interface, and the like.

  As shown in FIG. 1, the analysis apparatus A is provided with three-way valves 51, 52, 53, and 54. Each of these three-way valves 51, 52, 53, 54 has three connection ports (not shown), and the communication state and the cutoff state of these connection ports are independently controlled by the control unit 71. .

  The electrophoretic solution tank 11 is connected to the three-way valve 51 via the flow path 61. The purified water tank 12 and the cleaning liquid tank 13 are connected to the three-way valve 53 via flow paths 62 and 63. The dilution tank 25 is connected to the three-way valve 51 via a flow path 64 and is connected to the three-way valve 54 via a flow path 67. The three-way valve 51 is connected to the three-way valve 52 via the flow path 65. The three-way valve 53 is connected to the three-way valves 52 and 54 via the branch flow path 68.

  Separation channels 30 a, 30 b, 30 c, and 30 d are connected to the downstream side of the three-way valve 52 via a branch channel 66. Pinch valves 55a, 55b, 55c, and 55d are provided in portions of the branch channel 66 that are connected to the separation channels 30a, 30b, 30c, and 30d. The pinch valves 55a, 55b, 55c, and 55d are controlled to be opened and closed by the control unit 71, and allow and block the inflow to the separation channels 30a, 30b, 30c, and 30d. A manifold 57 is connected to the downstream side of the separation channels 30a, 30b, 30c, and 30d. Pinch valves 56a, 56b, 56c, and 56d are provided between the separation flow paths 30a, 30b, 30c, and 30d and the manifold 57. The pinch valves 56a, 56b, 56c, and 56d are controlled to be opened and closed by the control unit 71, and the communication state and the cutoff state of the separation flow paths 30a, 30b, 30c, and 30d and the manifold 57 are independently controlled.

  The manifold 57 is connected to the three-way valve 54 via a flow path 69. A waste liquid bottle 58 is connected to the downstream side of the three-way valve 54. The waste liquid bottle 58 is for storing a used liquid. A suction pump 59 is connected to the waste liquid bottle 58. The suction pump 59 generates a negative pressure. This negative pressure is applied to the branch flow path 68 and the flow path 69 via the three-way valve 54. The manifold 57 has a function of causing the negative pressure applied through the flow path 69 to act equally on the separation flow paths 30a, 30b, 30c, and 30d.

  The power supply unit 72 is for applying a voltage for performing analysis by capillary electrophoresis in the separation channels 30a, 30b, 30c, and 30d. As shown in FIG. 2, the power supply unit 72 is connected to the electrodes 33a, 33b, 33c, and 33d and the electrodes 34a, 34b, 34c, and 34d. Switches 73a, 73b, 73c, and 73d are provided between the power supply unit 72 and the electrodes 33a, 33b, 33c, and 33d. The switches 73a, 73b, 73c, and 73d are ON / OFF controlled by the control unit 71. Thereby, the power supply part 72 and the electrodes 33a, 33b, 33c, and 33d can be independently conducted. The voltage applied by the power supply unit 72 is, for example, about 1.5 kV. In the following description, the power supply unit 72 will be described as an example in which the electrodes 33a, 33b, 33c, and 33d are applied to the positive electrode, and the electrodes 34a, 34b, 34c, and 34d are applied to the negative electrode. You may provide the function to apply a voltage with the opposite polarity to this.

  Next, an analysis method using the analyzer A will be described below.

  FIG. 3 is a flow showing a process performed by using any one of the separation channels 30a, 30b, 30c, and 30d among the processes performed in the present analysis method. Hereinafter, a case where the separation channel 30a is used will be described as an example. This flow is roughly divided into a pretreatment step S1 and an analysis step S2. Pretreatment process S1 consists of washing process S11, filling process S12, and dispensing process S13.

  The cleaning step S11 is a step of cleaning the sample Sp and the like used in the previous analysis remaining in the separation channel 30a prior to the analysis step S2. Specifically, in FIG. 1, the three-way valve 53 is switched to a state where the three-way valve 53 communicates from the purified water tank 12 and the cleaning liquid tank 13 to the branch flow path 68 according to a command from the control unit 71. Further, the three-way valve 52 is switched to a state in which the three-way valve 52 communicates from the branch channel 68 to the branch channel 66. The pinch valves 55a and 56a are opened, and the pinch valves 55b, 55c, 55d, 56b, 56c, and 56d are closed. The three-way valve 54 is in a state of communicating from the branch flow path 68 to the waste liquid bottle 58. In this state, the suction pump 59 is operated by the control unit 71. Due to the negative pressure generated thereby, the purified water L2 and the cleaning liquid L3 are filled in the separation flow path 30a, and these are discharged to the waste liquid bottle 58. In addition, you may implement washing | cleaning process S11 by pouring purified water L2 separately after flowing the washing | cleaning liquid L3 to the separation flow path 30a.

  The filling step S12 is a step of filling the separation channel 30a with the electrophoresis liquid L1 for realizing electrophoresis. Specifically, the three-way valve 51 is switched to a state in which the flow path 61 and the flow path 65 communicate with each other and the flow path 64 is blocked by these in response to a command from the control unit 71. The three-way valve 52 is switched to a state in which the flow path 65 and the branch flow path 66 communicate with each other and the branch flow path 68 is blocked from these. The pinch valves 55a, 55b, 55c, 55d, 56a, 56b, 56c, 56d, and the three-way valve 54 are in the same state as the above-described cleaning step S11. In this state, the suction pump 59 is operated. Thereby, the electrophoretic liquid L1 is filled in the flow path 30a.

  The dispensing step S13 is a step of dispensing the sample Sp from the introduction hole 31a to the separation channel 30a. Further, the dispensing step of the present embodiment includes a step of diluting the sample Sp into a state suitable for analysis. Specifically, as shown in FIG. 4, the nozzle 22 is caused to penetrate the lid 24 by the drive mechanism (not shown) described above according to a command from the control unit 71. Then, the tip of the nozzle 22 is immersed in the sample Sp. Next, the sample Sp is extracted into the syringe 21 through the nozzle 22 by causing the syringe 21 to perform an inhalation operation.

  On the other hand, in FIG. 1, the three-way valve 51 is switched to a state where the flow path 61 and the flow path 64 communicate with each other. The electrophoretic liquid L1 is introduced into the dilution tank 25 by generating pressure using a pump (not shown). Next, as shown in FIG. 5, the nozzle 22 is caused to enter the electrophoresis liquid L <b> 1 in the dilution tank 25 by the drive mechanism. Then, the sample Sp is introduced into the dilution tank 25 by causing the syringe 21 to perform a discharging operation. At this time, in order to promote stirring of the sample Sp and the electrophoretic liquid L1, it is preferable to cause the syringe 21 to repeatedly perform an intake operation and an exhaust operation. In the present embodiment, the sample Sp is, for example, whole blood containing hemoglobin, and the electrophoretic solution L1 contains a hemolytic component that exhibits a hemolytic action that destroys the blood cell membrane. As a result, the blood cells of the sample Sp are hemolyzed and become a state suitable for hemoglobin analysis.

  Next, the sample Sp diluted in the dilution tank 25 is sucked by the syringe 21. As shown in FIG. 6, the tip of the nozzle 22 is caused to enter the introduction hole 31a of the separation channel 30a by the drive mechanism. In this state, by causing the syringe 21 to perform a discharging operation, the diluted sample Sp is introduced into the introduction hole 31a. As described above, the pretreatment step S1 is completed, and the analysis in the separation channel 30a is possible.

  In the present embodiment, the dispensing step S13 includes a dilution step. However, when the sample Sp that does not require dilution is an analysis target, the dispensing step S13 may be performed without going through the dilution step.

  Next, the analysis step S2 is performed. As shown in FIG. 3, the analysis step S2 includes a separation step S21 and a detection step S22.

  The separation step S21 is a step of separating, for example, hemoglobin, which is a specific component contained in the sample Sp, in the electrophoretic liquid L1 filled in the separation channel 30a. Specifically, in the circuit shown in FIG. 2, the switch 72 a is switched to the ON state according to a command from the control unit 71. Then, a voltage is applied from the power supply unit 72 to the electrodes 33a and 34a. At this time, the electrode 33a is a positive electrode and the electrode 34a is a negative electrode. As a result, an electroosmotic flow from the electrode 33a to the electrode 34a is generated in the electrophoretic liquid L1. In addition, hemoglobin, which is a specific component, moves in accordance with the inherent electrophoretic mobility. Since this moving speed differs depending on the substance, hemoglobin as a specific component moves from the electrode 33a toward the electrode 34a at a speed different from that of the other components.

  The detection step S22 is a step of detecting, for example, the amount or concentration of hemoglobin that is the separated specific component. Specifically, according to a command from the control unit 71, the detection unit 41a irradiates, for example, light having a wavelength of 415 nm from the above-described light source (not shown) to the part of the separation channel 30a that is marked with a circle in FIG. To do. Then, the reflected light is received by the light receiving unit described above. When hemoglobin, which is a specific component separated, passes through the portion irradiated with light from the light source unit, the absorbance grasped from the light receiving state of the light receiving unit changes. By processing this change by the control unit 71, the amount or concentration of hemoglobin can be detected. The detection result is stored in the memory of the control unit 71 as the analysis result of the separation channel 30a. From the above, the pretreatment step S1 and the analysis step S2 in the separation channel 30a are completed.

  The pretreatment step S1 and the analysis step S2 described above are the same when performed using the separation flow paths 30b, 30c, and 30d. By opening and closing the pinch valves 55a, 55b, 55c, 55d, 56a, 56b, 56c, and 56d, the separation channels 30a, 30b, 30c, and 30d can be implemented independently.

  Next, the pretreatment process S1 and the analysis process S2 using the separation channels 30a, 30b, 30c, and 30d will be described. As shown in FIG. 7, the pretreatment step S1a and the analysis step S2a are performed using the separation channel 30a, the pretreatment step S1b and the analysis step S2b are performed using the separation channel 30b, and the separation channel 30c is formed. The pretreatment step S1c and the analysis step S2c are used to perform the pretreatment step S1d and the analysis step S2d using the separation channel 30d. The preprocessing steps S1a, S1b, S1c, and S1d and the analysis steps S2a, S2b, S2c, and S2d are subscripted for convenience and are the same as the preprocessing step S1 and the analysis step S2, respectively. . Also, the cleaning steps S11a, S11b, S11c, S11d, filling steps S12a, S12b, S12c, S12d, dispensing steps S13a, S13b, S13c, S13d, and separation steps S21a, S21b, S21c, S21d, detection steps S22a, S22b, S22c and S22d are steps similar to the above-described washing step S11, filling step S12, dispensing step S13, separation step S21, and detection step S22, respectively.

  First, the pretreatment step S1a using the separation channel 30a is started. When the cleaning step S11a is completed, the pretreatment step S1a using the separation channel 30b is started. In parallel with this, the filling step S12a, the dispensing step S13a, and the analysis step S2a using the separation channel 30a are performed. When the cleaning step S11b using the separation channel 30b is completed, the pretreatment step S1c using the separation channel 30c is started. In parallel with this, the filling step S12b, the dispensing step S13b, and the analysis step S2b using the separation channel 30b are performed. When the cleaning step S11c using the separation channel 30c is completed, the pretreatment step S1d using the separation channel 30d is started. In parallel with this, the filling step S12c, the dispensing step S13c, and the analysis step S2c using the separation channel 30c are performed. When the cleaning step S11d is completed, the filling step S12d, the dispensing step S13d, and the analysis step S2d using the separation channel 30d are subsequently performed. Thus, in the present embodiment, the cleaning steps S11a, S11b, S11c, and S11d are continuously performed without delay. The preprocessing steps S1a, S1b, S1c, S1d and the analysis steps S2a, S2b, S2c, S2d are set at different start times.

  Next, the operation of the analyzer A and the analysis method using the same will be described.

  According to the present embodiment, as shown in FIG. 7, the time required to complete the preprocessing steps S1a, S1b, S1c, S1d is the time required to complete each of the preprocessing steps S1a, S1b, S1c, S1d. Obviously shorter than the total time. For this reason, it is possible to finish analysis process S2a, S2b, S2c, S2d considerably early compared with the example shown, for example in FIG. Thereby, the analysis using the analyzer A can be performed more efficiently.

  As shown in FIG. 1, the separation channels 30 a, 30 b, 30 c, and 30 d are connected to a common electrophoresis solution tank 11, purified water tank 21, and washing solution tank 13 via a branch channel 66. Since the pinch valves 55a, 55b, 55c, and 55d provided on the upstream side of the separation flow paths 30a, 30b, 30c, and 30d can be opened and closed individually, the separation liquid L1, the purified water L2, and the washing liquid L3 are separated from each other. It is possible to selectively flow into 30a, 30b, 30c, and 30d. This is suitable for carrying out the pretreatment steps S1a, S1b, S1c, S1d in parallel.

  As shown in FIG. 2, by providing the switches 73a, 73b, 73c, and 73d, it is possible to selectively apply a voltage from the common power supply unit 72 to the separation channels 30a, 30b, 30c, and 30d. . Furthermore, it is possible to apply a voltage to any two of the separation channels 30a, 30b, 30c, and 30d at the same time. This is suitable for performing the detection steps S22a, S22b, S22c, and S22d in parallel.

  Since the nozzle 22 of the dispensing means 20 can penetrate the lid 24, the sample Sp can be appropriately extracted from the sample container 23. In addition, by repeating the suction operation and the exhaust operation of the syringe 21, it is possible to favorably promote the stirring of the sample Sp in the dilution tank 25 and the electrophoretic liquid L1 as the diluent. The nozzle 22 can individually dispense the sample Sp into the introduction holes 31a, 31b, 31c, and 31d of the separation channels 30a, 30b, 30c, and 30d. As a result, the sample Sp can be dispensed without delay to one of the separation channels 30a, 30b, 30c, 30d in which one of the filling steps S12a, S12b, S12c, S12d has been completed.

  By providing the manifold 57, it is possible to suppress the negative pressure from the suction pump 59 from being applied unevenly to the separation flow paths 30a, 30b, 30c, and 30d.

  In addition, it is also possible to implement a method in which dispensing by the nozzle 22 is performed on the discharge holes 32a, 32b, 32c, and 32d. In this case, the power supply unit 72 applies a voltage so that the electrodes 34a, 34b, 34c, and 34d are positive and the electrodes 33a, 33b, 33c, and 33d are negative. Since the detection units 41a, 41b, 41c, and 41d are arranged at positions shifted from the center of the separation channels 30a, 30b, 30c, and 30d, detection is performed from the start point of movement by electrophoresis by reversing the polarity. The moving distances to the parts 41a, 41b, 41c, 41d will be different. Thereby, analysis under different conditions can be performed.

  The analysis apparatus and the analysis method according to the present invention are not limited to the above-described embodiments. The specific configuration of the analysis apparatus and analysis method according to the present invention can be varied in design in various ways.

  The configuration may be such that the analysis is performed on the same sample Sp in any two or more of the separation channels 30a, 30b, 30c, and 30d. In this case, the control unit 71 calculates the final analysis result of the sample Sp by averaging the analysis results obtained by the plurality of analyzes. In addition, when any of the analysis results performed a plurality of times deviates greatly from the range assumed in advance, or is far from the average value of the plurality of analysis results, the control unit 71 Such an analysis result may be determined as an abnormal result. Those determined to be abnormal analysis results are removed from the averaging process.

  It is good also as a structure which performs a correction | amendment calculation process by the control part 71 with respect to the analysis result for every separation flow path 30a, 30b, 30c, 30d. For example, when hemoglobin (Hb) is a measurement target, a calibration sample having a known concentration is obtained in each of the separation channels 30a, 30b, 30c, and 30d prior to the above-described pretreatment step S1 and analysis step S2. Analyze it. By comparing the analysis result (the ratio of HbA1c) obtained by this analysis with the known ratio of HbA1c of the calibration sample, the correction coefficient for each separation channel 30a, 30b, 30c, 30d is calculated. deep. Then, for each of the separation channels 30a, 30b, 30c, and 30d, the correction calculation process using the correction coefficient is performed on the analysis result obtained by the actual measurement. According to such a configuration, the analysis result can be corrected to a more accurate value.

  Another method of the correction calculation process is to correct the correlation between the moving speed and the measurement result. In analysis using electrophoresis, the slower the moving speed, the longer it takes to pass through the detection units 41a, 41b, 41c, and 41d. Even if the actual concentration is the same, the measured value tends to increase as the passage time increases. For this reason, it is good to prepare the correction coefficient according to the time which detection requires about each of separation flow path 30a, 30b, 30c, 30d. By performing correction calculation processing on the actual analysis result using the correction coefficient corresponding to the time required for detection, variation in the analysis result due to the moving speed can be suppressed. It is conceivable that the correlation between the detection time and the analysis result changes depending on the number of times the separation channels 30a, 30b, 30c, and 30d are used. For this reason, it is preferable to set the correction coefficient described above as a value having a correlation with the number of uses.

  The number of separation flow paths 30a, 30b, 30c, and 30d is not limited to four. The configuration of the separation channels 30a, 30b, 30c, and 30d is not limited to a so-called straight type, and may be, for example, a cross injection type in which two channels intersect. The sample Sp is not limited to one containing hemoglobin represented by whole blood, and may contain, for example, DNA, RNA (ribonucleic acid), or protein.

  The analysis performed in the analysis step in the present invention is not limited to the one using capillary electrophoresis, and may be one using a micro liquid chromatography method, for example. In this case, separation, elution, reaction and the like in the column are performed in the separation step, and the reaction purified product is detected in the detection step.

A Analyzing apparatus L1 Electrophoretic liquid L2 Purified water L3 Cleaning liquid Sp Sample 10 Storage tank 11 Electrophoretic liquid tank 12 Purified water tank 13 Cleaning liquid tank 20 Dispensing means 21 Syringe 22 Nozzle 23 Sample container 24 Lid 25 Dilution tanks 30a, 30b, 30c, 30d Separation Flow path 31a, 31b, 31c, 31d Inlet hole 32a, 32b, 32c, 32d Discharge hole 33a, 33b, 33c, 33d Electrode 34a, 34b, 34c, 34d Electrode 36a, 36b, 36c, 36d Main body 40 Detection means 41a, 41b , 41c, 41d Detectors 51, 52, 53, 54 Three-way valves 55a, 55b, 55c, 55d Pinch valves 56a, 56b, 56c, 56d Pinch valves 57 Manifold 58 Waste liquid bottle 59 Suction pump (pressure generating means)
61, 62, 63, 64, 65, 67, 69 Channel 66, 68 Branch channel 71 Control unit 72 Power supply unit 73a, 73b, 73c, 73d Switch

Claims (32)

A plurality of separation channels for separating a specific component contained in the introduced sample;
Detection means for detecting the separated specific component;
A control unit that controls a separation step for performing the separation in the separation channels and a detection step having a detection step for performing the detection, and a pretreatment step for setting the separation channels in a state where the analysis step can be performed When,
A dispensing means having a nozzle for dispensing the sample into the separation channels,
The pretreatment step further includes a dispensing step of dispensing the sample into the separation channel,
The separation step utilizes electrophoresis,
The plurality of separation channels are connected to a channel for filling an electrophoresis solution branched from one electrophoresis solution tank in which an electrophoresis solution used for electrophoresis is stored, and the nozzle is inserted, and An analyzer in which an introduction hole for introducing the sample is formed.   The analysis device according to claim 1, wherein the detection unit includes a plurality of detection units each provided in each separation channel. Each of the detection units performs detection at a position biased to either end from the center of each separation channel, and
The analyzer according to claim 2, wherein a sample can be introduced into each separation channel from both ends.   The analyzer according to any one of claims 1 to 3, wherein a pressure generating means for applying a pressure capable of discharging the liquid filled in each of the separation channels is connected to the plurality of separation channels.   The analyzer according to claim 4, wherein a manifold for uniformizing pressure applied to the plurality of separation channels is provided between the plurality of separation channels and the pressure generating unit. A sample container having a lid for containing the sample and blocking the sample from outside air;
The analyzer according to claim 1, wherein the nozzle can penetrate the lid. The pretreatment step includes a dilution step of diluting the sample with a diluent in a dilution tank,
The analyzer according to any one of claims 1 to 6, wherein the sample and the diluent are stirred by suction and discharge from the nozzle to the dilution tank.   The analyzer according to claim 7, wherein the electrophoresis solution also serves as the diluent. Each of the separation channels has a pair of electrodes provided near both ends,
A common power supply for applying a voltage capable of causing electrophoresis in each of the separation channels;
A switch capable of selecting which one of the pair of electrodes of the plurality of separation flow paths is electrically connected;
The analyzer according to claim 1, wherein the switch switching operation is controlled by the control unit.   The analyzer according to claim 9, wherein the power supply unit is capable of switching a polarity of a voltage applied to each separation channel.   11. The analyzer according to claim 8, wherein a cross section of each separation channel is a circle having a diameter of 25 to 100 μm or a rectangle having a side length of 25 to 100 μm.   The analyzer according to claim 1, wherein the sample includes hemoglobin.   The analyzer according to claim 12, wherein the sample is blood.   14. The control unit according to claim 1, wherein when the analysis step is performed in two or more separation channels for the same sample, the control unit averages the analysis results in the separation channels. The analyzer described in 1.   The analyzer according to claim 14, wherein the control unit performs the averaging process except for an analysis result of the two or more separation channels that is determined to be abnormal. The analyzer according to claim 14 or 15 , wherein the control unit performs a correction calculation process on the analysis result using a correction coefficient set for each of the separation channels. An analysis step including a separation step of separating a specific component of a sample introduced into each of the separation channels using a plurality of separation channels, and a detection step of detecting the separated specific component by a detection unit;
A pre-treatment step that makes each of the separation channels in a state where the analysis step can be performed,
The pretreatment step further includes a dispensing step of dispensing the sample into the separation channel using a dispensing means having a nozzle.
The separation step utilizes electrophoresis,
In the pretreatment step, before the dispensing step, the electrophoresis solution used for electrophoresis is separated from one electrophoresis solution tank in which the electrophoresis solution is stored and connected to the plurality of separation channels. A filling step of filling the separation channel;
In the dispensing step, an analysis method in which the nozzle is introduced into an introduction hole provided in the separation channel and the sample is introduced.   The analysis method according to claim 17, wherein the detection unit includes a plurality of detection units each provided in each separation channel. Each of the detection units performs detection at a position biased to either end from the center of each separation channel, and
The analysis method according to claim 18, wherein a sample can be introduced into each separation channel from both ends thereof.   The analysis method according to any one of claims 17 to 19, wherein a pressure generating means for applying a pressure capable of discharging the liquid filled in each of the separation channels is connected to the plurality of separation channels.   21. The analysis method according to claim 20, wherein a manifold for equalizing pressure applied to the plurality of separation channels is provided between the plurality of separation channels and the pressure generating unit.   The analysis according to any one of claims 17 to 21, wherein the sample is sucked by penetrating a lid provided in a sample container for containing the sample to shield the sample from outside air. Method. The pretreatment step includes a dilution step of diluting the sample with a diluent in a dilution tank,
The analysis method according to any one of claims 17 to 22, wherein the sample and the diluent are stirred by suction and discharge from the nozzle to the dilution tank.   The analysis method according to claim 23, wherein the electrophoresis solution also serves as the diluent. Each of the separation channels has a pair of electrodes provided near both ends,
The analysis method according to any one of claims 17 to 24, wherein a voltage capable of causing electrophoresis is selectively applied to the pair of electrodes of each separation channel by switching a switch from a common power supply unit. .   The analysis method according to claim 25, wherein the power supply unit is capable of switching a polarity of a voltage applied to each separation channel.   27. The analysis method according to claim 24, wherein a cross section of each separation channel is a circle having a diameter of 25 to 100 [mu] m or a rectangle having a side length of 25 to 100 [mu] m.   The analysis method according to any one of claims 17 to 27, wherein the sample includes hemoglobin.   The analysis method according to claim 28, wherein the sample is blood.   30. The analysis method according to any one of claims 17 to 29, wherein when the analysis step is performed in two or more separation channels for the same sample, the analysis results in the separation channels are averaged. .   The analysis method according to claim 30, wherein the averaging process is performed except for the analysis result of the two or more separation flow paths that is determined to be abnormal. 32. The analysis method according to claim 30 or 31, wherein correction calculation processing is performed on the analysis result using a correction coefficient set for each of the separation channels.

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