JP2008175798A - Cartridge for analysis, and absorbance measuring device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a cartridge for analysis suited to POCT with an LC method adopted therefor, that is, a simple and compact cartridge for analysis right fit for detecting glycohemoglobin with the LC method adopted therefor. <P>SOLUTION: The use of a separation column 40 with less pressure loss makes it possible to provide this cartridge 10 for analysis of small size with glycohemoglobin detection performance maintained. Further, this cartridge 10 includes an operation plate 31 which can be positioned at an initial position with a specimen holding hole 36 exposed to the exterior while it is not used yet and which can be positioned at an introduction position with the holding hole 36 forming part of an inflow passage 23 when it is used. Further, the operation plate 31 can be repositioned from the initial position to the introduction position with simple operation which is slide operation. This compact and simply operated cartridge 10 right fit for detecting glycohemoglobin can be thereby provided. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to an analytical cartridge used for analyzing a measurement sample and an absorbance measuring apparatus suitable for the same.

2. Description of the Related Art Conventionally, in hospital inspection facilities and inspection centers, blood is used as a measurement sample, and a specific component contained therein is measured using a liquid chromatography (hereinafter referred to as “LC”) device. For example, Patent Document 1 discloses an LC device that analyzes glycohemoglobin, which is an index for long-term control of diabetes. In this LC device, the eluent A and eluent B stored in two bottles are sucked by the feed pump, and sucked by the feed pump via the deaerator, the eluent switching valve, and the eluent mixing manifold. Is done. The mixed solution of the eluent A and the eluent B that has passed through the liquid feed pump is introduced into the separation column together with the blood sample introduced from the sample introduction unit. Then, glycohemoglobin is detected by a spectrophotometer arranged on the downstream side of the column.
JP 2001-74721 A

  LC measurement using an existing column requires a large apparatus because the back pressure in the column increases. This has hindered the use for point-of-care testing (POCT) performed by medical staff in the field of medical care and nursing. On the other hand, for example, DCA2000 manufactured by Bayer is known as an apparatus for detecting glycohemoglobin in consideration of POCT. However, since this apparatus employs a latex immunoagglutination method, it employs an LC method. Therefore, development of a small analytical cartridge suitable for POCT is desired.

  The present invention has been made in view of such problems, and an object thereof is to provide a small analytical cartridge suitable for POCT using the LC method. Another object is to provide an absorbance measurement apparatus capable of performing LC analysis using such an analytical cartridge.

  The present invention adopts the following means in order to achieve at least one of the above objects.

The analytical cartridge of the present invention is:
A first porous silica part having an average pore diameter of 0.5 to 5 μm and a cation-exchangeable first porous silica part and an average pore diameter of 1 to 5 μm and a hydrophobic second porous silica part A separation column in which the average pore diameter of the second porous silica part is different from the average pore diameter of the second porous silica part;
One or more cylinders for storing liquid to be supplied to the separation column;
An inflow path for guiding the liquid supplied from the supply port of the one or more cylinders to the inlet of the separation column;
An outflow path for guiding the liquid flowing out from the outlet of the separation column to the waste liquid port;
A detection passage provided to form a part of the outflow path and capable of detecting a component in the liquid by an external detection device;
It is equipped with.

In the analysis cartridge of the present invention, when the measurement sample is introduced into the inflow passage, the liquid stored in one or more cylinders is supplied to the entrance of the separation column together with the measurement sample, and the components contained in the measurement sample are separated by the separation column. Separated, flows out from the outlet of the separation column to the outflow passage, and component detection is performed by the external detection device in the detection passage provided in the outflow passage. The analytical cartridge has a first porous silica portion having an average pore diameter of 0.5 to 5 μm and a cation exchange property, and a second porous silica portion having an average pore diameter of 1 to 5 μm and a hydrophobic property, Since LC analysis is performed using a separation column in which the average pore diameter of the first porous silica portion and the average pore diameter of the second porous silica portion are different, the back pressure of the separation column can be sufficiently reduced. Therefore, the overall size can be reduced despite the adoption of the LC method. Therefore, it is possible to provide an analytical cartridge suitable for POCT employing the LC method, that is, a simple and small analytical cartridge suitable for detection of glycohemoglobin employing the LC method.

  In the analytical cartridge of the present invention, the first porous silica part may have an average pore diameter of 1 to 2.3 μm, and the second porous silica part may have an average pore diameter of 2 to 4.2 μm. . In this way, analysis can be performed with higher accuracy.

  In the analytical cartridge of the present invention, the first porous silica part is subjected to a surface treatment with a sulfonate, and the second porous silica part is made of a silane compound having an alkyl chain having 4 to 23 carbon atoms. Surface treatment may be performed. By doing so, it is possible to separate the three types of stable glycohemoglobin, unstable glycohemoglobin, and hemoglobin in blood more accurately. Here, examples of the sulfonate include 3-sulfopropyl potassium methacrylate, and examples of the silane compound include octadecyldimethylchlorosilane.

  In the analysis cartridge according to the present invention, the inflow path may have at least one or more U-shaped curved portions. In this way, the liquid that has flowed out from the supply port of each cylinder can be mixed at a shorter distance than when the inflow path is a straight line, so that the entire analysis cartridge can be further downsized.

  In the analysis cartridge according to the present invention, only one cylinder is provided and divided into a plurality of chambers by a plurality of movable partition walls slidable in a liquid-tight manner, and liquids having different compositions are stored in the plurality of chambers. In use, when the most distal movable partition is pressed toward the tip of the cylinder, the movable partition on the distal side among the plurality of movable partitions sequentially reaches the distal end surface of the cylinder, and each time it enters the supply port. The facing room may be configured to change. In this case, it is not necessary to store liquids having different compositions in a plurality of cylinders and mix the liquids in a desired ratio, and thus the entire analysis cartridge can be further downsized.

  The analysis cartridge of the present invention includes supply port sealing means for sealing each supply port of the cylinder so as to be openable and closable, and a liquid is sealed in the cylinder by the supply port sealing means. Also good. In this way, the trouble of preparing the liquid at the time of measurement can be saved, so that the trouble of the operator can be reduced. At this time, the supply port sealing means may be a single sealing plate that can open and close the supply ports of the cylinder all at once by a sliding operation. If it carries out like this, since each supply port can be open | released all at once, an operator's effort can be reduced more.

The analysis cartridge according to the present invention includes an outflow passage sealing unit that seals a portion of the outflow passage other than the detection passage so as to be openable and closable, and liquid is supplied to the separation column and the outflow passage. It is good also as what is enclosed with the sealing means. In this way, since the deterioration of the separation column can be prevented by prefilling the separation column with the solution, a highly accurate analytical cartridge can be provided. The liquid sealed by the supply port sealing means and the outflow passage sealing means may have the same composition as that of one kind of liquid stored in advance in each cylinder, or may have a different composition. . At this time, the supply port sealing means and the outflow passage sealing means may be a common plate that can open and close the supply ports and the outflow passage of the cylinder at the same time by a sliding operation. In this way, since all the sealing means can be opened at the same time, the labor of the operator can be reduced.

  The analysis cartridge of the present invention includes an outflow passage sealing means for sealing the portion other than the detection passage of the outflow passage so as to be openable and closable. When there are two or more cylinders, the supply port of one cylinder is always opened and the supply ports of the remaining cylinders are opened and closed so that they can be opened and closed. The opened cylinder and the separation column may be filled with liquid by the outflow passage sealing means, and the remaining cylinder may be filled with liquid by the supply port sealing means. In this way, since the separation column can be filled with the liquid stored in advance in the cylinder, the deterioration of the separation column can be prevented, and even when a plurality of cylinders are provided, the liquid in each cylinder is mixed. Can be prevented. At this time, the supply port sealing means may be a single sealing plate that can simultaneously open the supply ports of the cylinder by a sliding operation. In this way, since a plurality of supply ports can be opened simultaneously, the labor of the operator can be reduced. Alternatively, the supply port sealing means and the outflow passage sealing means may be a common plate that can open the supply ports and the outflow passage of the cylinder all at once by a sliding operation. In this way, all the sealing means can be opened at the same time, so that the labor of the operator can be further reduced.

  In the analysis cartridge according to the present invention, the supply port sealing means may be a pressure valve that operates when the pressure of the liquid reaches a predetermined pressure to open each supply port of the cylinder. In this way, the supply port can be opened without bothering the operator.

  In the analysis cartridge according to the present invention, the outflow passage sealing means may be a pressure valve that operates when the pressure of the liquid reaches a predetermined pressure to open the outflow passage. In this way, the outflow passage can be opened without bothering the operator.

  The analysis cartridge according to the present invention includes an operation plate having a sample holding hole for holding a measurement sample, and the operation plate is positioned at an initial position where the sample holding hole is arranged outside when not in use. Sometimes, a part of the inflow path may be formed by being positioned at an introduction position where the measurement sample can be introduced into the separation column from the sample holding hole. At this time, the operation plate may be positioned from the initial position to the introduction position by a slide operation. In this way, the measurement sample can be supplied to the separation column by a simple operation of sliding the operation plate from the initial position to the introduction position.

  The analysis cartridge of the present invention may be provided with a sample holding position confirmation means that is provided close to the sample holding hole, is exposed to the outside when the sample is not added, and is covered with the added sample when the sample is added. If it carries out like this, it can be confirmed by the sample holding position confirmation means whether the sample is appropriately hold | maintained at the sample holding hole.

  The analysis cartridge according to the present invention includes an introduction amount adjusting unit that removes an excess sample when the operation plate moves from the initial position to the introduction position while the sample is held in the sample holding hole. Also good. In this way, the amount of the measurement sample supplied to the separation column can be made constant by a simple operation of switching the operation plate from the initial position to the introduction position. At this time, the introduction amount adjusting unit scrapes off the sample existing outside the sample holding hole when the operation plate moves from the initial position to the introduction position among the samples held in the sample holding hole. It may be a thing.

  The analysis cartridge of the present invention includes a dilution tank for storing a liquid for diluting the measurement sample, and the operation plate can be positioned at an intermediate position between the initial position and the introduction position. The sample holding hole may be provided in the dilution tank when the operation plate is positioned at the intermediate position. In this way, since the measurement sample can be diluted to a concentration suitable for measurement and supplied to the separation column, the labor for adjusting the concentration of the sample in advance can be saved. As a specific method of dilution, manual vibration agitation may be performed in a state where the sample holding hole is disposed inside the dilution tank, or vibration agitation using an ultrasonic vibrator may be performed.

  The analysis cartridge of the present invention may include a drop-off prevention mechanism that prevents the operation plate from dropping off. By so doing, it is possible to prevent the operation plate from falling off and to prevent the liquid in the cartridge from leaking to the outside.

  The analysis cartridge of the present invention may include a waste liquid tank that receives the liquid flowing out from the outlet of the separation column through the waste liquid port. In this way, it is not necessary to prepare a waste liquid tank separately from the analysis cartridge. In this case, the waste liquid tank may be a waste liquid tank having an exhaust hole with a hydrophobic film that exhausts air in the waste liquid tank. In this way, it is possible to prevent only the gas from being discharged and the liquid in the waste liquid tank from being discharged to the outside. The one or more cylinders may be accommodated in the waste liquid tank. In this way, the entire analysis cartridge can be reduced in size compared to the case where the waste liquid tank and one or more cylinders are provided separately.

  The layout of the cylinder and separation column in the analysis cartridge of the present invention is not limited to the above-described form, and the analysis cartridge of the present invention may be discarded after being used once for analysis of the measurement sample. Good.

The absorbance measuring apparatus of the present invention is
An installation section on which any of the cartridges for analysis described above can be installed;
A pressing mechanism capable of independently pressing the liquid in the one or more cylinders of the analysis cartridge installed in the installation unit;
A light-emitting element disposed on one side of the detection path and a light-receiving element disposed on the other side of the detection path are configured to allow light emitted from the light-emitting element to enter the detection path and pass through the detection path. A detector that receives the subsequent light at the light receiving element;
It is equipped with.

  In this absorbance measurement apparatus, first, an analysis cartridge is installed in the installation section. When this analytical cartridge is installed in the installation section of the absorbance measuring device, it is possible to press one or more cylinders by the pressing mechanism. At this time, the liquid composition supplied to the separation column can be set to a desired composition by individually adjusting the pressure applied to the cylinder. When the cylinder is pressed, the liquid stored in the cylinder is supplied to the separation column together with the measurement sample, the components contained in the measurement sample flow out to the outflow passage while being separated by the separation column, and the detection provided in the outflow passage Absorbance can be measured by detecting the liquid passing through the passage with a detector. Thus, according to the absorbance measuring apparatus of the present invention, LC analysis using any of the above-described analytical cartridges can be easily performed.

  The pressing mechanism may be composed of, for example, a piston rod inserted into the cylinder and an actuator (for example, a motor, an air pressure cylinder, or a hydraulic cylinder) that moves the piston rod in the axial direction of the cylinder. Moreover, the detector may be comprised by LED as a light emitting element, and the phototransistor as a light receiving element.

  In the absorbance measuring apparatus of the present invention, the detector includes at least one of a diffusion plate that diffuses light of the light emitting element or a polarizing plate that suppresses the influence of reflected light between the light emitting element and the detection path. It may be configured. When a diffuser plate is used, the illuminance distribution can be made uniform even when, for example, a light emitting LED having a luminance spot is adopted. Moreover, when a polarizing plate is used, the influence of reflected light or the like can be suppressed. By doing so, absorbance measurement can be accurately performed in any case.

  Next, embodiments of the present invention will be described with reference to the drawings. FIG. 1 is a front view of an analysis cartridge 10 according to an embodiment of the present invention, FIG. 2 is a plan view of the analysis cartridge 10, and FIG. 3 is a sectional view of the analysis cartridge taken along the line AA in FIG. 4 is an explanatory view showing the removal of blood held outside the sample holding hole 36, and FIG. 5 is an explanatory view for explaining the movement of the first closing valve 26a. is there. The analysis cartridge 10 of the present embodiment supplies a substantially rectangular body 20, first and second cylinders 21 and 22 arranged substantially in parallel inside the body 20, and an appropriate amount of sample to the separation column 40. An operation plate 31, a separation column 40 for separating the measurement sample, and a waste liquid tank 50 for storing the liquid that has passed through the separation column 40. The analysis cartridge 10 is intended for diagnosis of diabetes, and when measuring the absorbance of each of the stable glycated hemoglobin, unstable glycated hemoglobin and hemoglobin separated from blood as a measurement sample. It is used for. The analysis cartridge 10 is a so-called disposable type, and is discarded after being used once for analysis of a measurement sample. Further, the entire size of the analysis cartridge 10 is about 20 mm in width (W), about 55 mm in depth (D), and about 10 mm in height (H).

The body 20 is made of a transparent acrylic resin, and as shown in FIG. 2, the inflow passage 23 connecting the first cylinder 21 and the second cylinder 22 and the inlet of the separation column 40, the outlet of the separation column 40, and the waste liquid tank. 50, an outlet passage 25 that connects to 50, a sample adjustment recess 33 that adjusts the amount of the measurement sample introduced into the separation column 40, a dilution tank 34 that dilutes the measurement sample, and a first supply port 21 b of the first cylinder 21. Other than the detection passage (here, the first closing valve 26a that seals the second supply port 22b of the second cylinder 22 and the second closing valve 26b that seals the second supply port 22b of the second cylinder 22). And a third closing valve 26c that seals the vicinity of the waste liquid port 51 in an openable and closable manner. The inflow passage 23 supplies the eluent A in the first cylinder 21 and the eluent B in the second cylinder 22 to the inlet of the separation column 40 via the first supply port 21b and the second supply port 22b. The inflow passage 23 has a solution mixing unit 27 at a position closer to the separation column 40 than a position where the eluent A and the eluent B merge. The eluent A and the eluent B are mixed in the solution mixing unit 27 which is a U-shaped curved portion, and the mixed solution is supplied to the inlet of the separation column 40. The outflow passage 25 discharges the liquid that has passed through the separation column 40 to the waste liquid tank 50 through the waste liquid port 51. A part of the outflow passage 25 serves as a detection passage 24, and a component in the liquid can be detected by an absorbance measurement device 60 which is an external detection device described later. The sample adjustment recess 33 is provided on the side surface of the body 20, and a slit-like operation plate insertion port 32 is provided on the bottom surface of the sample adjustment recess 33. The operation plate 31 is inserted in close contact with the operation plate insertion port 32 (see FIG. 4A). The dilution tank 34 stores a diluted solution, and the sample held in the sample holding hole 36 of the operation plate 31 is diluted with the operation of the operation plate 31 described later. As shown in FIG. 5, the first closing valve 26 a is attached to the first supply port 21 b so that a cylindrical member 110 whose inside is hollowed out is orthogonal, and is rotatable inside the cylindrical member 110. The rotating member 112 is in close contact. A linear notch 116 is provided on the upper surface of the rotating member 112, and a through hole 114 is provided on the side surface. The first closing valve 26a is in a sealed state where the through-hole 114 is not connected to the communication port 118 provided in the columnar member 110 (see FIG. 5A), and in an open state where it is connected to the communication port 118 (FIG. 5). (B) can be selected. Similarly, the second closed valve 26b and the third closed valve 26c can be selected from a sealed state and an open state. When not in use, all the shut-off valves are in a sealed state and the inflow passage 2
3. The eluent A is liquid-tightly sealed in the separation column 40 and the outflow passage 25 by a first closing valve 26a, a second closing valve 26b, and a third closing valve 26c. The liquid filled at this time may be another liquid.

  As shown in FIG. 2, the first cylinder 21 and the second cylinder 22 are arranged substantially in parallel inside the body 20. The first cylinder 21 is connected to the inflow passage 23 connected to the inlet of the separation column 40 through the first supply port 21b, and the second cylinder 22 is connected to the separation column 40 through the second supply port 22b. 23. Since the first closing valve 26a and the second closing valve 26b are in a sealed state when not in use, the eluent A is liquid-tightly sealed in the first cylinder 21 by the first closing valve 26a and the first rubber packing 21a. The eluent B is liquid-tightly sealed in the second cylinder 22 by the second closing valve 26b and the second rubber packing 22a. The eluent A and the eluent B are liquids having different salt concentrations.

  As shown in FIG. 2, the operation plate 31 includes a sample holding hole 36 and is inserted in close contact with the operation plate insertion port 32. The operation plate 31 has a surface made of a hydrophobic material, and the sample holding hole 36 has a hydrophilic property so that the measurement sample can be easily held by coating with a hydrophilic material. In addition, a reference line 37 is provided around the sample holding hole 36 (see FIG. 1), and it can be confirmed whether or not the sample is held in the sample holding hole 36. Further, the operation plate 31 has an initial position (see FIG. 3A) where the sample holding hole 36 is disposed outside the body 20 and an introduction position where the sample holding hole 36 forms a part of the inflow passage 23 (FIG. 3). (See (d)) and an intermediate position (see FIG. 3C) between the initial position and the introduction position and where the sample holding hole 36 is located in the dilution tank 34. When the operation plate 31 is operated according to this positioning, a sample having a concentration suitable for measurement can be supplied to the separation column 40 by a simple operation of pushing the operation plate 31 into the body 20.

  The separation column 40 is a polymer-bonded column in which the surface of the porous silica continuous carrier is chemically modified by a polymerization reaction. The surface of the porous silica continuous carrier is subjected to sol-gel transition and then calcined to obtain mesopores. Is a porous silica continuous carrier support column. The separation column 40 is composed of a cation exchange silica monolith disk (hereinafter referred to as “disk M”) and a hydrophobic silica monolith disk (hereinafter referred to as “disk O”), and the average pore diameter of the disk M is 0.5 to 5 μm. The average pore diameter of the disk O is preferably 1 to 2.3 μm, and the average pore diameter of the disk O is preferably 1 to 5 μm, and more preferably 2 to 4.2 μm. At this time, the disk M is a silica monolith disk surface treated with a sulfonate, and examples of the sulfonate include 4-vinylphenyl sulfonic acid and 3-sulfopropyl potassium methacrylate. Of these, potassium 3-sulfopropyl methacrylate is preferred. The disk O is preferably surface-treated with a silane compound having an alkyl chain having 4 to 23 carbon atoms. Examples of the silane compound include octyltrichlorosilane, octadecyltrichlorosilane, octyldimethylchlorosilane, and octadecyl. Examples thereof include dimethylchlorosilane, among which octadecyldimethylchlorosilane is preferable. The separation column 40 is a stack of a plurality of disks O and disks M produced by the above-described method. As a combination of the disk M and the disk O, the disk M is disposed in the upstream portion in the liquid feeding direction. It is preferable that the disk O is disposed in the downstream portion in the liquid feeding direction. Furthermore, the separation column 40 may be aged with bovine serum albumin. In this way, for example, when analyzing a blood component, it is possible to prevent the component to be analyzed from being adsorbed on the column. Also, since bovine serum albumin has a different absorbance from glycated hemoglobin and hemoglobin, even if bovine serum albumin is mixed in the purified sample, there is not much trouble in analyzing glycated hemoglobin or hemoglobin in the blood. .

  As shown in FIG. 2, the waste liquid tank 50 is a substantially rectangular parallelepiped space hollowed out from the body 20, and an outflow passage connected to the outlet of the separation column 40 via a waste liquid port 51 provided at an end of the waste liquid tank 50. 25, which receives the waste liquid that has passed through the separation column 40, and is filled with air when not in use. An exhaust hole 52 is provided on the upper surface of the waste liquid tank 50. Since the exhaust hole 52 is covered with a hydrophobic film 53 that is a hydrophobic porous film, gas can pass but liquid cannot pass. An operation plate discharge port 54 into which the operation plate 31 is inserted is provided on the side surface of the waste liquid tank 50, and the operation plate 31 is moved from the initial position (see FIG. 3A) to the introduction position (see FIG. 3D). ), The end of the operation plate 31 is pushed into the waste liquid tank 50. Further, since the drop-off prevention unit 38 provided at the end of the operation plate 31 is larger than the operation plate discharge port 54, the operation plate 31 is prevented from dropping from the operation plate insertion port 32, and the liquid in the cartridge is prevented. Leakage can be prevented. Further, since the operation plate 31 has a length that is positioned at the introduction position when it contacts the inner wall of the waste liquid tank 50, the measurement sample is separated from the separation column 40 by a simple method of sliding the operation plate 31 to the innermost position. Can be supplied to.

  Next, the operation of the analysis cartridge 10 of the present embodiment configured as described above will be described. 3A is a part of a cross-sectional view of the analysis cartridge 10 when not in use, cut along the AA plane in FIG. 1, and FIG. 3D shows the analysis cartridge 10 in use in FIG. It is a part of sectional drawing cut | disconnected by the AA surface. First, a description will be given of the case where the analysis cartridge 10 is not in use, with reference to FIG. When the analysis cartridge 10 is not used, the operation plate 31 is positioned at the initial position (see FIG. 3A). A patient who undergoes a simple test for diabetes causes a thin needle to lightly stab a fingertip to cause slight bleeding, and then presses the bleeding finger against the entrance of the sample holding hole 36 to introduce blood into the sample holding hole 36. At this time, the operator can confirm that the sample holding hole 36 is filled with blood by visually observing whether or not the reference line 37 is hidden by blood. Further, since the sample holding hole 36 is coated with a hydrophilic material, blood can be held without a gap.

  Subsequently, an operation when the blood filled in the sample holding hole 36 is introduced into the inflow passage 23 will be described. After visually confirming that the sample holding hole 36 is filled with blood, the operator pushes the operation plate 31 in a direction indicated by an arrow in FIG. Here, when the excess blood 37a is held in the sample holding hole 36, when the operation plate 31 is moved to the position shown in FIG. Adjusted to

  Here, the mechanism by which the excess sample is removed will be described in detail with reference to FIG. FIG. 4 is an explanatory view showing the removal of blood held outside the sample holding hole 36. As shown in FIG. 4A, the excess blood 37 a may be held outside the sample holding hole 36. If the excess blood 37a is held outside the sample holding hole 36, there is a possibility that a measurement error may occur due to an excessive introduction amount. However, in this embodiment, when the operation plate 31 is pushed in, the operation plate 31 is sized to be in close contact with the operation plate insertion port 32. Therefore, as shown in FIG. Surplus blood 37 a held outside the hole 36 is scraped off by the outer wall of the operation plate insertion port 32. Thereby, the amount of blood held in the sample holding hole 36 can be made constant. Further, the excess blood 37a scraped off at this time remains in the sample adjustment recess 33, so that it is possible to prevent the surrounding blood from being contaminated by the removed blood.

Subsequently, when the operator further pushes the operation plate 31 into the body 20 and is positioned at the intermediate position as shown in FIG. 3C, the sample holding hole 36 is located inside the dilution tank 34. Since the diluted solution is stored in the dilution tank 34 in advance, the sample holding hole 36 is used.
The blood introduced into can be diluted to a concentration suitable for measurement. Specifically, manual amplitude agitation is performed in a state where the sample holding hole 36 is disposed inside the dilution tank 34. In this way, the blood and the diluted solution can be mixed uniformly in a shorter time. The dilution tank 34 may be filled with the diluted solution, but it is preferable to mix air and the diluted solution in consideration of efficiently diluting the blood in the sample holding hole 36. At this time, since the sample holding hole 36 is coated with a hydrophilic material, there is no possibility that air enters the sample holding hole 36. In addition, since the analysis cartridge 10 is made of a transparent acrylic resin, the operator can visually check the degree of dilution by the color of the liquid in the dilution tank 34.

  Subsequently, when the operator further pushes the operation plate 31 into the body 20 and is positioned at the introduction position as shown in FIG. 3D, the sample holding hole 36 forms a part of the inflow passage 23. At this time, the dropout prevention portion 38, which is the end portion of the operation plate 31, is in contact with the inner wall of the waste liquid tank 50. Therefore, the operator can easily position the sample holding hole 36 at a position that becomes a part of the inflow passage 23 by inserting the operation plate 31 to the innermost position, and the measurement sample held in the sample holding hole 36. Can be supplied to the separation column 40.

  Subsequently, the operator changes the first closing valve 26a, the second closing valve 26b, and the third closing valve 26c from the sealed state (see FIG. 5A) to the opened state (see FIG. 5B). Thus, it becomes possible to supply blood having a concentration suitable for measurement to the separation column 40. This state is referred to as a use state of the analysis cartridge 10. Here, selection of the sealed state and the open state will be described in detail with reference to FIG. 5, taking the first closing valve 26a as an example. FIG. 5 is an explanatory diagram for explaining the operation of the first closing valve 26a. When the first closing valve 26a is in the sealed state, as shown in FIG. 5A, the through-hole 114 is in a position not connected to the inflow passage 23. Can not flow. On the other hand, in the open state, as shown in FIG. 5B, the through-hole 114 is connected to the inflow passage 23 via the communication port 118 and forms part of the inflow passage 23, so that the liquid flows. Can do. Selection between the sealed state and the open state is performed by inserting a flathead screwdriver or the like into a linear notch 116 attached to the upper portion of the rotating member 112 and rotating the rotating member 112. The same applies to the opening operation of the second closing valve 26b and the third closing valve 26c.

  After the analysis cartridge 10 is switched to the use state, the analysis cartridge 10 is set in the cartridge installation part 60 a of the absorbance measuring device 60. FIG. 6 is a schematic configuration diagram of an absorbance measuring device 60 in which the analysis cartridge 10 is set. The absorbance measuring device 60 moves the first piston rod 61a by the first stepping motor 61, thereby pressing the eluent A in the first cylinder 21 through the first rubber packing 21a, and independently of this, By moving the second piston rod 62a by the two-stepping motor 62, the eluent B in the second cylinder 22 is pressed through the second rubber packing 22a. A controller 63 incorporating a known microcomputer drives and controls the first stepping motor 61 and the second stepping motor 62 to adjust the pressing force of the eluent A and the pressing force of the eluent B. The liquid composition supplied to can be made a desired composition. The eluent A and the eluent B pressed by the first cylinder 21 and the second cylinder 22 are mixed through the solution mixing unit 27 of the inflow passage 23 and supplied to the separation column 40. The liquid that has passed through the separation column 40 passes through the detection passage 24 provided in the outflow passage 25 and is discharged to the waste liquid tank 50 through the waste liquid port 51.

Of the light from the LED light source 64 that is a light emitting element, the light that has passed through the dichroic mirror 76 enters the detection path 24 through the first lens 65, the diffusion plate 66, and the polarizing plate 67, and then passes through the detection path 24. Then, the measurement phototransistor 69 as a light receiving element is entered through the second lens 68. On the other hand, the light reflected by the dichroic mirror 76 enters the luminance adjustment phototransistor 78. Light entering the measurement phototransistor 69 and the brightness adjustment phototransistor 78 is detected by the detection circuit 70, and the analog signal after detection is converted into a digital signal by the A / D conversion circuit 72 and input to the controller 63. The controller 63 obtains the luminance based on the digital signal obtained from the luminance adjustment phototransistor 78 via the detection circuit 70 and the A / D conversion circuit 72, and the LED light source 64 has a uniform luminance. Feedback control of current. In addition, the controller 63 controls the lighting circuit 74 so that the LED light source 64 is repeatedly turned on and off, and converts the digital signal obtained from the measurement phototransistor 29 via the detection circuit 70 and the A / D conversion circuit 72. Based on the difference between when the LED light source 64 is turned on and when the LED light source 64 is turned off, a graph representing the relationship between the LC retention time and the absorbance is output to a display or printer (not shown). By performing such synchronous detection, absorbance measurement can be performed not only in a dark box but also under a fluorescent lamp.

  Here, the correspondence between the components of the present embodiment and the present invention will be clarified. The analysis cartridge 10 of the present embodiment corresponds to an analysis cartridge of the present invention, the separation column 40 corresponds to a separation column, the first cylinder 21 and the second cylinder 22 correspond to one or more cylinders, The first supply port 21b and the second supply port 22b correspond to supply ports, the inflow passage 23 corresponds to an inflow passage, the waste liquid port 51 corresponds to a waste liquid port, the outflow passage 25 corresponds to an outflow passage, and the detection passage 24 Corresponds to the detection passage, the solution mixing portion 27 corresponds to the U-shaped curved portion, the first closing valve 26a and the second closing valve 26b correspond to the supply port sealing means, and the third closing valve 26c flows out. Corresponding to the passage sealing means, the operation plate 31 corresponds to the operation plate, the sample holding hole 36 corresponds to the sample holding hole, the reference line 37 corresponds to the sample holding position confirmation means, and the operation plate insertion port 32 is introduced. Corresponds to the amount adjustment part, The release tank 34 corresponds to a dilution tank, the drop prevention part 38 and the operation plate discharge port 54 correspond to a drop prevention mechanism, the waste liquid tank 50 corresponds to a waste liquid tank, the exhaust hole 52 corresponds to an exhaust hole, and is hydrophobic. The membrane 53 corresponds to a hydrophobic membrane, the absorbance measurement device 60 corresponds to an absorbance measurement device, the cartridge installation portion 60a corresponds to an installation portion, and the first stepping motor 61 and the second stepping motor 62 correspond to a pressing mechanism. The LED light source 64 corresponds to a light emitting element, the measurement phototransistor 69 corresponds to a light receiving element, the diffusion plate 66 corresponds to a diffusion plate, and the polarizing plate 67 corresponds to a polarizing plate.

  According to the analysis cartridge 10 of this embodiment described in detail above, when blood is introduced into the inflow passage 23, the eluents A and B stored in the first cylinder 21 and the second cylinder 22 are separated from the separation column 40 together with the blood. The components supplied to the inlet and contained in the blood are separated by the separation column 40, flow out from the outlet of the separation column 40 to the outflow passage 25, and the components are detected by the absorbance measurement device 60 in the detection passage 24 provided in the outflow passage 25. Detection is performed. Since this analysis cartridge 10 performs LC analysis using the separation column 40, the back pressure of the separation column 40 can be sufficiently lowered. Can be small. Therefore, it is possible to provide an analytical cartridge suitable for POCT employing the LC method, that is, a simple and small analytical cartridge suitable for detection of glycohemoglobin employing the LC method.

  Further, the disk M of the separation column 40 is surface-treated with 3-sulfopropyl potassium methacrylate, which is a sulfonate, and the disk O is octadecyldimethyl, which is a silane compound having an alkyl chain having 4 to 23 carbon atoms. Since the surface treatment with chlorosilane is performed, three types of stable glycohemoglobin, unstable glycohemoglobin, and hemoglobin in blood can be separated with higher accuracy. Here, examples of the sulfonate include 3-sulfopropyl potassium methacrylate, and examples of the silane compound include octadecyldimethylchlorosilane.

Furthermore, by providing a solution mixing part 27 in the inflow passage 23 of the analysis cartridge 10, the first
Since the eluent A and the eluent B flowing out from the supply ports of the cylinder 21 and the second cylinder 22 can be mixed at a shorter distance than when the inflow passage 23 is a straight line, the entire analysis cartridge 10 is more It can be downsized.

  Furthermore, the first and second cylinders 21a and 26b are provided to seal the supply ports of the first cylinder 21 and the second cylinder 22 of the analysis cartridge 10 so that they can be opened and closed. 22, since the eluent A and the eluent B are sealed by the first closing valve 26a and the second closing valve 26b, it is possible to save the trouble of preparing the liquid at the time of measurement. Can be reduced.

  In addition, a third closing valve 26c is provided that seals the portion of the outflow passage 25 of the analysis cartridge 10 other than the detection passage 24 so that it can be opened and closed. Since it is sealed by the shutoff valve 26c, the separation column 40 can be prevented from deteriorating by prefilling the separation column 40 with the eluent A, so that a highly accurate analytical cartridge can be provided. it can.

  Further, the analysis cartridge 10 includes an operation plate 31 having a sample holding hole 36 for holding blood. The operation plate 31 is positioned at an initial position where the sample holding hole 36 is arranged outside when not in use. In use, the sample holding hole 36 forms a part of the inflow passage 23 by being positioned at an introduction position where the measurement sample can be introduced into the separation column 40 from the sample holding hole 36. At this time, since the operation plate 31 is positioned from the initial position to the introduction position by the slide mechanism, the operation plate 31 is positioned from the initial position to the introduction position by a simple operation of sliding the operation plate 31. Can do.

  Furthermore, since the guide line 37 is provided close to the sample holding hole 36 and exposed to the outside when blood is not added, and is covered with the added blood when blood is added, whether the reference line 37 is exposed or not. Therefore, blood can be appropriately held in the sample holding hole 36, and a more accurate measurement result can be obtained.

  In addition, the analysis cartridge 10 includes an operation plate insertion port 32 for removing excess blood 37a when the operation plate 31 moves from the initial position to the introduction position while holding the blood in the sample holding hole 36. Since the operation plate insertion port 32 scrapes off the excess blood 37a existing outside the sample holding hole 36, the separation column 40 can be easily operated by switching the operation plate 31 from the initial position to the introduction position. The amount of blood supplied to can be made constant.

  On the other hand, the analysis cartridge 10 includes a dilution tank 34 for storing a diluted solution for diluting blood, and the operation plate 31 can be positioned at an intermediate position between the initial position and the introduction position. Since the sample holding hole 36 is provided inside the dilution tank 34 when the plate 31 is positioned at the intermediate position, the blood is diluted to a concentration suitable for measurement and supplied to the separation column 40. This saves the effort of adjusting the blood concentration in advance. Further, as a specific method of dilution, manual vibration agitation is performed in a state where the sample holding hole 36 is disposed inside the dilution tank 34. However, vibration agitation using an ultrasonic vibrator or the like may be performed. Good.

  On the other hand, since the analysis cartridge 10 includes the drop-off prevention unit 38 that prevents the operation plate 31 from falling off, the operation plate 31 is prevented from falling off and the liquid in the analysis cartridge 10 is prevented from leaking to the outside. be able to.

On the other hand, the analysis cartridge 10 allows the liquid flowing out from the outlet of the separation column 40 to flow into the waste liquid outlet 5.
1 is provided, it is not necessary to prepare a waste liquid tank separately from the analysis cartridge 10. At this time, since the waste liquid tank 50 includes the exhaust hole 52 with the hydrophobic film 53 that exhausts the air in the waste liquid tank 50, only the gas is discharged and the liquid in the waste liquid tank 50 is discharged to the outside. Can be prevented.

  On the other hand, the controller 63 of the absorbance measuring device 60 controls the pressing force with which the first stepping motor 61 and the second stepping motor 62 press the first piston rod 61a and the second piston rod 62a individually, so that the separation column 40 can be controlled. Since the liquid composition to be supplied can be set to a desired composition, LC analysis using the analysis cartridge 10 can be easily performed. In addition, since the diffusion plate 66 and the polarizing plate 67 are provided between the LED light source 64 and the detection passage 24, changes in the incidence rate and the emission rate to the detection passage 24 are reduced, and stable measurement is possible.

  It should be noted that the present invention is not limited to the above-described embodiment, and it goes without saying that the present invention can be implemented in various modes as long as it belongs to the technical scope of the present invention.

In the above-described embodiment, the first closing valve 26a, the second closing valve 26b, and the third closing valve 26c are provided. However, the first closing valve 26a may be omitted. In this case, the liquid in the first cylinder 21 is transferred to the first cylinder 21 and the inflow passage 2 by the second closing valve 26b and the third closing valve 26c.
3. It is enclosed in the separation column 40 and the outflow passage 25. Even without the first closing valve 26a, the separation column 40 can be filled with the eluent A in advance, so that the separation column 40 can be prevented from being deteriorated and a highly accurate analysis cartridge can be provided. Further, since the second cylinder 22 is sealed by the second closing valve 26b, there is no possibility that the eluent A and the eluent B are mixed.

  In the embodiment described above, the inflow passage 23 is opened and closed by rotating the first closing valve 26a. However, as shown in FIG. 7, a pressure valve 100 may be provided. In this way, the outflow passage can be opened without bothering the operator. The outer periphery of the pressure valve 100 includes a deformable pressure valve support portion 102 and a pressure valve destruction portion 104 that is destroyed by pressure. Here, the movement of opening the pressure valve 100 will be described in detail with reference to FIG. FIG. 7 is an explanatory diagram for explaining the movement of the pressure valve 100. As shown in FIG. 7A, the pressure valve 100 at the time of sealing is blocked by the pressure valve 100 because the pressure valve support portion 102 and the pressure valve destruction portion 104 are in close contact with the inflow passage 23. And the liquid cannot flow. On the other hand, when the pressure valve 100 is released, as shown in FIG. 7B, the pressure valve destruction part 104 is destroyed by pressure, and the pressure valve support part 102 is deformed in the liquid traveling direction, so that the liquid flows. It will be ready. At this time, since the pressure valve destruction part 104 has a structure thinner than the other part of the pressure valve, when pressure is applied to the pressure valve 100 from the side, the pressure valve destruction part 104 is destroyed. . Further, since the pressure valve support portion 102 can be deformed, the pressure valve 100 can be prevented from coming off from the inflow passage 23. Although the first closing valve 26a is replaced with the pressure valve 100 here, it goes without saying that the same effect can be obtained by replacing the second closing valve 26b and the third closing valve 26c.

  In the embodiment described above, the first cylinder 21 and the second cylinder 22 are provided in the body 20, but may be provided in the waste liquid tank 50 as shown in FIG. In this case, the entire analysis cartridge 10 can be downsized as compared with the case where the waste liquid tank 50 and the first cylinder 21 and the second cylinder 22 are separately provided.

In the above-described embodiment, the operator opens the first closing valve 26a, the second closing valve 26b, and the third closing valve 26c so that blood having a concentration suitable for measurement is supplied to the separation column 40. As shown in FIG. 9, blood may be supplied to the separation column 40 by sliding the sealing plate 130 held by the plate holder 132. The plate holder 132 is made of soft rubber such as silicone rubber, and is formed by bonding two wall members so that a gap is formed. The gap serves as a slit for holding the sealing plate 130. As shown in FIG. 10, the two wall members constituting the plate holder 132 are provided with a first cylinder communication hole 134 that communicates the first supply port 21 b of the first cylinder 21 and the inflow passage 23, and the second cylinder 22. A second cylinder communication hole 136 for communicating the second supply port 22b and the inflow passage 23 and a waste liquid port communication hole 138 for communicating the waste liquid port 51 of the waste liquid tank 50 and the outflow passage 25 are provided in this order. . The sealing plate 130 is made of a synthetic resin such as polyester or polyethylene, and the first through hole 135 that communicates with the first cylinder communication hole 134 and the second through hole 137 that communicates with the second cylinder communication hole 136 by an operator's operation. A third through hole 139 that communicates with the waste liquid port communication hole 138 is provided. Further, since the plate holder 132 is made of a hydrophobic hard rubber, the liquid tightness is maintained at the connection portion of each hole. Here, the inner surface of each hole of the plate holder 132 is preferably hydrophilic, but may be hydrophobic. By doing so, the operator can make the analysis cartridge 10 usable more simply because the operator slides the sealing plate 130 to open them all at once. Further, the sealing plate 130 may only open and close the inflow passage 23. By doing so, even if the analysis cartridge 10 does not have the third closing valve 26c, the analysis cartridge 10 can be put into a usable state by sliding the sealing plate 130.

  In the embodiment described above, separate eluent A and eluent B are sealed in the first cylinder 21 and the second cylinder 22, but as shown in FIG. 11, the first movable partition 121, the second movable partition A cylinder 120 divided into a plurality of chambers 120x, 120y, and 120z is prepared by 122 and rubber packing 123, and separate eluent X, eluent Y, and eluent Z are stored in each of the chambers 120x, 120y, and 120z. You may make it do. Then, after the operator opens the fourth closing valve 127 and the third closing valve 26c, and sets the analysis cartridge in the absorbance measuring device 60, the rubber packing 123 of the cylinder 120 is directed toward the tip of the cylinder 120 by the piston rod. And press. Then, as shown in FIG. 11, the eluent X stored in the first chamber 120x is supplied from the supply port 120a to the entrance of the separation column 40, and then, as shown in FIG. When the first movable partition 121 reaches the front end surface of the cylinder 120, the eluent Y stored in the second chamber 120y is now supplied from the supply port 120a to the inlet of the separation column 40, and thereafter, FIG. As shown in FIG. 12, when the second movable partition wall 122 reaches the front end surface of the cylinder 120, the eluent Z stored in the third chamber 120z is supplied from the supply port 120a to the inlet of the separation column 40, as shown in FIG. As shown in (c), when the rubber packing 123 reaches the front end surface of the cylinder 120, the cylinder 120 becomes empty. In this way, the chamber connected to the supply port 120a is switched each time when it reaches the front end surface of the cylinder 120 in order from the movable partition wall on the front end side, and different eluents are supplied to the inlet of the separation column 40. There is no need to store liquids of different compositions in the cylinder and mix each liquid in the desired ratio. In addition, a relief groove 125 is formed between the inlet 124 of the passage from the inside of the cylinder 120 to the supply port 120a and the front end surface of the cylinder 120. After the first movable partition wall 121 exceeds the inlet 124, the eluent X is discharged. It is guided to the entrance 124 through the escape groove 125. This also applies to the eluent Y. Instead of the fourth closing valve 127 and the third closing valve 26c, a sealing plate that can open the inflow passage 23 and the outflow passage 25 at the same time is provided, and the sealing plate is operated by sliding operation. The analysis cartridge 10 may be in a usable state.

  In the embodiment described above, the body 20 includes the dilution tank 34, but the dilution tank 34 may be omitted. In this way, the entire analysis cartridge 10 can be further downsized.

  In the analysis cartridge of the above-described embodiment, the solution mixing unit 27 is provided in the inflow passage 23 to uniformly mix the two types of eluent. However, the porous filter that facilitates mixing of the two types of eluent May be disposed in the middle of the inflow passage 23 or immediately before the entrance of the separation column 40. In this way, the inflow passage 23 can be further shortened, and the analysis cartridge 10 can be further downsized.

  In the embodiment described above, the waste liquid tank 50 is built in the analysis cartridge 10. However, the waste liquid tank 50 may be externally attached to the analysis cartridge 10. In this way, the analysis cartridge 10 can be further downsized.

  In the above-described embodiment, the size of the analysis cartridge 50 is a very compact size having a width (W) of about 20 mm, a depth (D) of about 55 mm, and a height (H) of about 10 mm. For example, it may be twice or three times larger than this, or may be larger. Further, the type, size, and eluent of the separation column 40 to be used may be appropriately determined according to the measurement sample.

  In the above-described embodiment, the periphery of the sample holding hole 36 is surrounded by the reference line 37. However, any position may be used as long as the position is close to the sample holding hole 36 and the position hidden by the blood is described. Thus, a member that changes color may be used.

  Next, this invention is demonstrated based on an Example. The separation column is a polymer-bonded column in which the surface of the porous silica continuous carrier is chemically modified by a polymerization reaction. After the sol-gel transition of the surface of the porous silica continuous carrier, firing is performed. A porous silica continuous support column with reduced mesopores was used. This separation column consists of a cation exchange silica monolith disc (hereinafter referred to as “disc M”) having an average pore size of 2.0 μm and a hydrophobic silica monolith disc (hereinafter referred to as “disc O”) having an average pore size of 3.0 μm. The average pore diameter of each disk is Shimadzu Autopore 9520 as an analyzer, and about 0.1 g of sample is collected in a standard cell for large pieces, and the mercury intrusion method is performed under the condition of an initial pressure of about 2 kpa. And measured. The thickness of each disk is 2 mm, and the inner diameter of the disk is 3.2 mm. Further, the disk M was subjected to surface treatment by introducing 3-methacryloxypropyltrimethoxysilane (MOP) as a spacer on the surface of the silica monolith disk, and then benzoyl peroxide as an initiator to methacrylic acid-3-sulfo. Propyl potassium (MASK) was added in an amount equivalent to 10 equivalents of silanol groups present on the disk surface, reacted for 24 hours under methanol reflux, washed with water and methanol, and then modified. Furthermore, the disk O removes water from the silica monolith disk, adds octadecyldimethylchlorosilane (ODS) and pyridine to 5 equivalents of silanol groups present on the disk surface, and reacts under toluene reflux for 24 hours. Then, it was washed with toluene, tetrahydrofuran (THF) and ethanol, and dried under reduced pressure to make ODS. This was prepared by reacting hexamethyldisilazane (HMDS) with ODS-modified silica monodisk for 17 hours under toluene reflux, washing with toluene, THF, and ethanol, and modifying. The disk O and the disk M manufactured by the above-described method are stacked in the order of “MMMOOO (M: disk M, O: disk O)”, filled in a heat shrinkable tube, and then contracted at about 220 degrees to form a separation column. Produced.

Table 1 below shows the results of measurement using the separation column prepared by the above-described method 10 times at each pressure loss and measuring how many times the leakage frequency is. The pressure loss was measured using a digital pressure sensor AP-V80 (manufactured by KEYENCE) with the pressure loss before and after the column set to 0 to 100 kgf / cm ² . As a result, as shown in Table 1 below, the pressure loss was 4
(Kgf / cm ² ) or less has no leakage, and since a column generally used in POCT is used at 5 (kgf / cm ² ), this separation column is sufficient for use in POCT. It can be said that it has performance.

  In this measurement, the separation column prepared by the method described above was connected to an HPLC apparatus, and eluent A (20 mM MES, 20 mM HEPES, 0.01 wt% sodium azide (pH 5.2)) and eluent B (20 mM MES, After passing through a mixed solution of 20 mM HEPES, 0.01 wt% sodium azide, 400 mM NaCl (pH 7.0)) mixed at A: B = 82: 18, 1% (w / v) bovine serum albumin aqueous solution Was injected 10 times continuously at a rate of 2 μl / time per disk O, and once fed under the conditions shown in Table 2 below, the sample was aged and then fixed to an analysis cartridge.

  In this measurement, an HPLC apparatus (manufactured by Shiseido, NANOSPACESI), pump (NANOSPACESI-2 3101), UV-VIS detector (NANOSPACESI-2 3002), degasser (NANOSPACESI-2 3010), column oven (NANOSPACESI-2 3014), A manual injector (NANOSPACESI-2 3006) and a data processing device (Shiseido S-MicroChrom) were used.

  Here, the result of having carried out the separation measurement of glycohemoglobin using this separation column is shown. The measurement was performed using an actual sample standard for HbA1c measurement (level 2: HbA1c is 5.38% (w / w)). JDS HbA1c Lot. 2 (manufactured by HECTEF Standard Reference Center, a limited liability intermediate corporation) diluted with ultrapure water 101 times, D-(+) glucose (manufactured by Sigma Aldrich Japan, special grade) at 10 mg / ml and 100 mg / ml ultrapure water The actual sample standard for HbA1c measurement was diluted 101 times with this glucose solution, and 2 μl of each incubated at 37 ° C. for 2 hours was added, and the conditions were as shown in Table 2 above. As a result, as shown in FIG. 13, stable HbA1c (outflow time around 10 minutes) can be separated from unstable HbA1c (outflow time around 8 minutes) and hemoglobin HbA (outflow time around 14 minutes). It was. For this reason, it can be said that the separation column prepared by the method described above has sufficient performance for detecting stable HbA1c.

  In addition, when the disk M and the disk O created by the above-described method were stacked in the combinations shown in Table 3 below and the same experiment was performed, as shown in FIGS. Could be detected. For this reason, it can be said that even when the combination of disks shown in Table 3 below is used, the disk has sufficient performance for detecting the stable HbA1c.

FIG. 3 is a front view of the analysis cartridge 10. FIG. 3 is a plan view of the analysis cartridge 10. FIG. 2 is a part of a cross-sectional view of the analysis cartridge taken along plane AA in FIG. An explanatory view showing the removal of blood held outside the sample holding hole. Explanatory drawing for demonstrating operation | movement of the 1st closing valve 26a. The schematic block diagram of the light absorbency measuring apparatus 60 which set the cartridge 10 for analysis. Explanatory drawing for demonstrating the motion of the pressure valve. The top view of the cartridge 10 for analysis of other embodiment (the 1). The top view of the cartridge 10 for analysis of other embodiment (the 2). Explanatory drawing for demonstrating opening and closing of the sealing plate 130. FIG. The top view of the cartridge 10 for analysis of other embodiment (the 3). Explanatory drawing for demonstrating a motion of the cartridge 10 for analysis of other embodiment (the 3). Chromatogram measured with a combination of “MMMOOO”. Chromatogram measured with a combination of “MOOMOMOOMO”. Chromatogram measured with a combination of “MMMMOOOO”. Chromatogram measured with a combination of “MOMOMOMO”. Chromatogram measured with a combination of “MMOOMMMO”.

Explanation of symbols

  10 analysis cartridge, 20 body, 21 first cylinder, 21a first rubber packing, 21b first supply port, 22 second cylinder, 22a second rubber packing, 22b second supply port, 23 inflow passage, 24 detection passage, 25 Outflow passage, 26a 1st closing valve, 26b 2nd closing valve, 26c 3rd closing valve, 27 solution mixing part, 31 operation plate, 32 operation plate insertion port, 33 sample adjustment recess, 34 dilution tank, 36 sample holding Hole, 37 Reference line, 37a Surplus blood, 38 Drop prevention part, 40 Separation column, 50 Waste liquid tank, 51 Waste liquid port, 52 Exhaust hole, 53 Hydrophobic membrane, 54 Operation plate discharge port, 60 Absorbance measuring device, 60a Cartridge installation Part, 61 first stepping motor, 61a first piston rod, 62 second stepping 62a second piston rod, 63 controller, 64 LED light source, 65 first lens, 66 diffuser plate, 67 polarizing plate, 68 second lens, 69 phototransistor for measurement, 70 detection circuit, 72 A / D conversion circuit , 74 lighting circuit, 76 dichroic mirror, 78 brightness adjusting phototransistor, 100 pressure valve, 102 pressure valve support part, 104 pressure valve breaking part, 110 columnar member, 112 rotating member, 114 through-hole, 116 notch, 118 Communication port, 120 cylinder, 120a Supply port, 120x, 120y, 120z Multiple chambers, 121 First movable partition, 122 Second movable partition, 123 Rubber packing, 124 Entrance, 125 Relief groove, 127 Fourth closing valve, 130 Seal Stop plate, 132 Plate holder, 134 No. 1 cylinder communication hole, 135 first through hole, 136 second cylinder communication hole, 137 second through hole, 138 waste liquid port communication hole, 139 third through hole.

Claims (26)

A first porous silica part having an average pore diameter of 0.5 to 5 μm and a cation-exchangeable first porous silica part and an average pore diameter of 1 to 5 μm and a hydrophobic second porous silica part A separation column in which the average pore diameter of the second porous silica part is different from the average pore diameter of the second porous silica part;
One or more cylinders for storing liquid to be supplied to the separation column;
An inflow path for guiding the liquid supplied from the supply port of the one or more cylinders to the inlet of the separation column;
An outflow path for guiding the liquid flowing out from the outlet of the separation column to the waste liquid port;
A detection passage provided to form a part of the outflow path and capable of detecting a component in the liquid by an external detection device;
Cartridge for analysis. The first porous silica part has an average pore diameter of 1 to 2.3 μm, and the second porous silica part has an average pore diameter of 2 to 4.2 μm.
The analysis cartridge according to claim 1. The first porous silica part is subjected to a surface treatment with a sulfonate, and the second porous silica part is subjected to a surface treatment with a silane compound having an alkyl chain having 4 to 23 carbon atoms.
The analysis cartridge according to claim 1 or 2. The inflow path has at least one or more U-shaped curved portions,
The analysis cartridge according to claim 1. The cylinder is provided with only one cylinder, and is divided into a plurality of chambers by a plurality of movable partition walls slidable in a liquid-tight manner. In the plurality of chambers, liquids having different compositions are stored, and the most movable at the time of use. When the partition wall is pressed toward the tip of the cylinder, the movable partition wall on the tip side among the plurality of movable partition walls sequentially reaches the tip surface of the cylinder, and the room facing the supply port changes each time. Being
The analysis cartridge according to claim 1. The analysis cartridge according to any one of claims 1 to 5,
Supply port sealing means for sealing each supply port of the cylinder so as to be openable and closable,
With
Liquid is sealed in the cylinder by the supply port sealing means,
Cartridge for analysis. The supply port sealing means is a single sealing plate that can simultaneously open and close the supply ports of the cylinder by a sliding operation.
The analysis cartridge according to claim 6. The analysis cartridge according to claim 6 or 7,
Outflow passage sealing means for sealing the portion other than the detection passage of the outflow passage so as to be openable and closable;
With
Liquid is sealed in the separation column by the supply port sealing means and the outflow passage sealing means,
Cartridge for analysis. The supply port sealing means and the outflow passage sealing means are one common plate capable of opening and closing each supply port and the outflow passage of the cylinder all at once by a slide operation.
The analysis cartridge according to claim 8. The analysis cartridge according to any one of claims 1 to 5,
Outflow passage sealing means for sealing the portion other than the detection passage of the outflow passage so as to be openable and closable;
With
When one cylinder is provided, the cylinder supply port is always opened. When two or more cylinders are provided, one cylinder supply port is always opened, and the remaining cylinder supply ports are opened. Having a supply port sealing means for sealing so as to be opened and closed;
Liquid is sealed in the normally open cylinder and the separation column by the outflow passage sealing means, and liquid is sealed in the remaining cylinders by the supply port sealing means,
Cartridge for analysis. The supply port sealing means is a single sealing plate that can simultaneously open the supply ports of the cylinder by a sliding operation.
The cartridge for analysis according to claim 10. The supply port sealing means and the outflow passage sealing means are one common plate capable of opening each supply port and the outflow passage of the cylinder all at once by a slide operation.
The cartridge for analysis according to claim 10. The supply port sealing means is a pressure valve that operates when the pressure of the liquid reaches a predetermined pressure and opens each supply port of the cylinder.
The analysis cartridge according to claim 6, 8 or 10. The outflow passage sealing means is a pressure valve that operates when the pressure of the liquid reaches a predetermined pressure to open the outflow passage.
The analysis cartridge according to claim 8, 10 or 13. The analysis cartridge according to any one of claims 1 to 14,
An operation plate having a sample holding hole for holding a measurement sample,
With
The operation plate is positioned at an initial position where the sample holding hole is arranged outside when not in use, and is positioned at an introduction position where the sample holding hole can be introduced into the separation column when used. Forming,
Cartridge for analysis. The operation plate is positioned from the initial position to the introduction position by a slide operation.
The analysis cartridge according to claim 15. The analysis cartridge according to claim 15 or 16,
A sample holding position confirmation means that is provided close to the sample holding hole, exposed to the outside when the sample is not added, and covered with the added sample when the sample is added;
Cartridge for analysis. The analysis cartridge according to any one of claims 15 to 17,
An introduction amount adjusting unit that removes surplus sample when moving from the initial position to the introduction position in a state where the operation plate holds the sample in the sample holding hole;
Cartridge for analysis. The introduction amount adjusting unit scrapes off a sample existing outside the sample holding hole when the operation plate moves from the initial position to the introduction position among the samples held in the sample holding hole. is there,
The analysis cartridge according to claim 18. The analysis cartridge according to any one of claims 15 to 19, comprising:
A dilution tank for storing a liquid for diluting the measurement sample;
With
The operation plate can be positioned at an intermediate position between an initial position and an introduction position;
The dilution tank is provided so that the sample holding hole is positioned inside the dilution tank when the operation plate is positioned at the intermediate position.
Cartridge for analysis. The analysis cartridge according to any one of claims 15 to 20, comprising:
A drop prevention mechanism for preventing the operation plate from falling off,
Cartridge for analysis. The analysis cartridge according to any one of claims 1 to 21,
A waste liquid tank for receiving the liquid flowing out from the outlet of the separation column through the waste liquid port;
Cartridge for analysis. The cartridge for analysis according to claim 22,
The waste liquid tank is a waste liquid tank having an exhaust hole with a hydrophobic film that exhausts air in the waste liquid tank.
Cartridge for analysis. The one or more cylinders are accommodated in the waste liquid tank.
The cartridge for analysis according to claim 22 or 23. An installation part capable of installing the analysis cartridge according to any one of claims 1 to 24;
A pressing mechanism capable of independently pressing the liquid in the one or more cylinders of the analysis cartridge installed in the installation unit;
A light-emitting element disposed on one side of the detection path and a light-receiving element disposed on the other side of the detection path are configured to allow light emitted from the light-emitting element to enter the detection path and pass through the detection path. A detector that receives the subsequent light at the light receiving element;
Absorbance measuring apparatus. The detector includes at least one of a diffusion plate that diffuses light of the light emitting element or a polarizing plate that suppresses the influence of reflected light between the light emitting element and the detection path.
The absorbance measurement apparatus according to claim 25.

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