JP2007155398A - Concentration element and chemical analyzer using it - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a concentration element capable of concentrating a liquid sample in a system without using a concentration element separately installed outside, and a chemical analyzer equipped with the concentration element. <P>SOLUTION: The concentration element is equipped with the flow channel formed on a substrate, the solvent separation layer installed in contact with at least a part of the side surface of the flow channel and the solvent holding layer installed through the solvent separation layer. The chemical analyzer is constituted so as to perform chemical analysis using the liquid sample and equipped with the concentration element and an introducing means for introducing the liquid sample into the concentration element. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

  The present invention relates to a concentration element for concentrating a liquid sample, and a chemical analyzer using the concentration element. In particular, the present invention can be suitably applied to a micro chemical analysis system (μTAS: Micro Total Analysis System) that performs chemical analysis or chemical synthesis on a chip.

  In recent years, research on μTAS has become active. μTAS is an abbreviation for Micro Total Analysis System, and is a micro chemical analysis system in which pumps, valves, sensors, etc. are miniaturized and integrated on a substrate such as glass or plastic.

  Advantages of this μTAS include reduction of sample amount and power consumption, reduction of reaction time, downsizing and disposability of equipment, low cost, and automation of measurement including pretreatment.

  The μTAS is considered to be able to exert its advantages particularly in the fields of medical care and environmental measurement, and is expected to be applied to, for example, an inexpensive disposable biosensor.

  However, there are not many fields where this μTAS is put into practical use. Therefore, in order to expand the application range, further improvement in analysis accuracy and reduction in analysis time are desired.

  The concentration of an analysis target component (hereinafter referred to as analysis component) in a liquid sample greatly affects analysis accuracy and analysis time in chemical analysis.

  For example, when the concentration of the analysis component in the liquid sample is very low, a physical or chemical change appears or it takes time until it becomes steady. In addition, the amount of change that appears is small, making detection difficult.

  In such a case, the sample needs to be concentrated to a certain concentration as a pretreatment for analyzing the sample.

  As a general concentration method, a method of separating and purifying analytical components using an ion exchange reaction, a method of extracting analytical components with an organic solvent, a method of separating a solvent using centrifugal force, and evaporating the solvent Various methods, such as a method of separating by the method, are used.

  These methods require extensive equipment and operation. Therefore, when the chemical analysis is performed with μTAS, whose miniaturization is the miniaturization of the apparatus, it is necessary to separately perform these concentration processes outside the system.

  However, when these concentration processes are performed outside the system, there are problems such as troublesome work, an increase in the total analysis time including pre-processing, and an extra analysis cost.

  Furthermore, when the sample after concentration treatment is introduced into μTAS, errors due to handling or contamination due to contamination with impurities may occur, which may affect analysis accuracy.

  In such a background, Japanese Patent Application Laid-Open No. 2004-053371 discloses that a liquid sample is heated by a heating element to evaporate the solvent for the purpose of concentrating the liquid sample without using an external concentration device. A chemical analysis method for concentrating the solution is disclosed.

JP-A-2005-31041 discloses a microreactor having a concentrating part for separating and adsorbing components in a liquid on a single substrate and an extracting part for extracting the components in the liquid by solvent. It is disclosed.
JP 2004-053371 A JP 2005-31041 A

  However, in order to further utilize the advantages of μTAS such as small size, low price and low power consumption, a simpler and cheaper concentration method is desired.

  Further, in the method disclosed in Japanese Patent Application Laid-Open No. 2004-053371, since the sample is heated and concentrated, the sample may be denatured if the heat resistance of the sample is low. For example, the heat resistance of biological materials used for biosensors is often relatively low, and this denaturation may be a problem.

  The method disclosed in JP-A-2005-31041 is a method in which a target component is adsorbed in a concentrating part, and then the target component is dissolved in an eluent to obtain a concentrated liquid.

  Therefore, since two processes of adsorption and dissolution are required, there remain problems that time is required and troublesome.

  Furthermore, components that cannot be adsorbed in the adsorption treatment and components that cannot be completely dissolved in the dissolution treatment are generated, and there may be many components that cannot be used in practice.

  The present invention has been made in view of the above problems, and an object of the present invention is to provide a simple concentration element and a chemical analyzer that can concentrate a sample in a μTAS system.

  In order to achieve the above object, the present invention provides a concentration element for concentrating a liquid sample, and a solvent separation installed in contact with a flow path formed on a substrate and at least a part of a side surface of the flow path. And a solvent holding layer installed via the solvent separation layer.

  The solvent separation layer is preferably made of a porous material, and the solvent retention layer is preferably provided with a material that absorbs the solvent.

  The flow path includes an inlet for introducing a liquid sample, and the cross-sectional area of the inlet is smaller than the cross-sectional area of the flow path. It is further desirable to have a configuration in which the discharge port is provided, and the cross-sectional area of the discharge port is smaller than the cross-sectional area of the introduction port, or the flow path includes a pressure loss portion.

  In addition, the present invention is a chemical analysis apparatus that performs a chemical analysis using a liquid sample, and includes the concentration element and an introduction unit that introduces the liquid sample into the concentration element.

  Furthermore, it is preferable to include a detection unit that detects the presence or absence or amount of a substance in the liquid sample.

  According to the present invention, a liquid sample can be concentrated in a μTAS system, and a simple concentration element and a chemical analyzer can be provided.

  A concentration element according to the present invention is a concentration element for concentrating a liquid sample, a channel formed on a substrate, a solvent separation layer installed in contact with at least a part of a side surface of the channel, And a solvent holding layer installed via a solvent separation layer. When the liquid sample is introduced into the flow path, the solvent contained in the liquid sample selectively passes through the solvent separation layer and is held in the solvent holding layer due to the pressure of the liquid. Therefore, the solvent is selectively removed from the liquid sample, and as a result, the liquid sample can be concentrated.

  Hereinafter, the present invention will be described in detail.

(Substrate)
The concentration element according to the present invention is suitably used for μTAS. Therefore, it is possible to use a substrate generally used for μTAS.

  Illustrative examples include inorganic materials such as glass, organic materials such as resin, semiconductor materials such as silicon, metal materials, and the like, and it is possible to form a flow path to be described later and has resistance to a liquid sample. This is not a limitation.

  Further, the substrate itself may also serve as a solvent separation layer and a solvent holding layer which will be described later.

(Flow path)
FIG. 1 is a diagram schematically showing an example of the shape of a flow path provided in the concentration element according to the present invention.

  The channel (3) in the present invention means a space into which the liquid sample (1) is introduced, and has at least an introduction port (2) for introducing the liquid sample.

  Furthermore, you may provide the discharge port (4) which discharges | emits a liquid sample (1) (FIG. 1 (a)). Then, in the flow path (3) provided in the concentration element in the present invention, the connection flow path (5) for introducing or discharging the liquid sample (1) has an introduction port (2) and a discharge port (4). It may be further connected via.

  Note that, as shown in FIG. 1B, the cross-sectional area of the inlet port (2) may be smaller than the cross-sectional area of the flow path (3). With such a configuration, it takes a long time for the liquid sample (1) to stay in the concentration element, and the concentration effect can be improved.

  Further, as shown in FIG. 1 (c), the cross-sectional area of the discharge port (4) may be smaller than the cross-sectional area of the introduction port (2). When the liquid sample flows through the concentration element, the amount of liquid is reduced by removing the solvent. As a result, the flow rate decreases. Therefore, when it is desired to reduce the change in flow velocity, the configuration as shown in FIG. 1 (c) is effective.

  Furthermore, a pressure loss part (6) may be provided in the flow path (3). Since the pressure loss part becomes a resistance obstacle to the flow of the liquid sample (1), it is possible to increase the concentration effect.

  In addition, the pressure loss part (6) may be provided by installing a structure as an aperture as shown in FIG. 1 (d), or the affinity of the inner wall surface of the flow channel with the liquid sample (1). Other methods may be used, such as providing a region with reduced properties (e.g., hydrophilicity when the liquid sample is an aqueous solution) (e.g., a water-repellent region). What is necessary is just to provide.

  These shapes are not limited to the shape in the plane direction, and may be configured in the depth direction (vertical direction with respect to the flow path). Further, the positions of the introduction port (2), the discharge port (4), and the connection channel (5) are not limited to the position configuration in the planar direction.

  For example, the connection channel (5) in FIG. 1 (b) may be formed in the longitudinal direction and connected to the channel (3) of the concentration element.

  The width of the flow path can be appropriately selected from the dimensions of about 1 μm to several mm, more preferably 1 μm to several 100 μm in consideration of the fluid control technique used in so-called μTAS.

  And by using such a fine flow path, it becomes possible to take much surface area with respect to the volume of the solution introduce | transduced. Therefore, the ratio of the area in contact with the solvent separation layer to the volume is increased, and the concentration effect can be enhanced.

  These flow paths are formed on the substrate. FIG. 2 is a diagram schematically illustrating an example of a cross section of the formed flow path. The flow path (3) may be formed on the base (10) as shown in FIG. 2 (a), but the first base (11) having the groove (14) as shown in FIG. It may be formed by superimposing the second substrate (12) to be the lid, and such a method is convenient and preferable in terms of the production method.

  The groove (14) may be formed by partially removing the substrate (13) by etching, cutting, or the like as shown in FIG. 3 (a), or as shown in FIG. 3 (b). (15) Another member (16) may be installed on the top and formed using a gap. Further, the substrate (13, 15) and the separate member (16) can also serve as a solvent separation layer and a solvent holding layer, which will be described later.

(Solvent separation layer)
The solvent separation layer is disposed in contact with at least a part of the side surface of the channel. The solvent separation layer in the present invention means a layer having a function of separating a solvent from a liquid sample. That is, the speed at which the analysis component permeates the membrane and the speed at which the solvent permeates the membrane are greatly different, and as a result, it means a layer through which the solvent mainly passes from the liquid sample.

  Therefore, it is desirable that the solvent separation layer has a function of selectively allowing the solvent to pass therethrough, but even if the passing solvent contains the analysis component, the solvent separation layer has a solvent content relative to the analysis component as compared with the liquid sample. If the ratio of the amounts is high, the effect of the present invention can be obtained. Note that the solvent may contain components other than the analysis component and may be separated at the same time.

  The separation utilizes the difference between the properties of the analysis component and the properties of the other components.

  For example, when there is a difference in the size of the analysis component molecule and the solvent molecule, separation is possible by using a porous material for the separation layer. If the pore size of the porous material is set to a size that allows solvent molecules to pass through but does not allow analysis components to pass through, the solvent mainly passes through this separation layer.

  As the porous material, a material used as a so-called semipermeable membrane, such as a cellulose membrane or a polyamide membrane, is preferably used. Many such semipermeable membranes having a separation ability corresponding to the molecular weight of various substances are commercially available, and are preferably used in the present invention.

  Further, inorganic materials such as mesoporous silica, porous silicon, and porous ceramics, and organic porous materials formed using phase separation can also be used.

  Furthermore, the porous material in the present invention includes not only a material having so-called pores but also a material in which pores are formed by bundling fine particles, fibers, hollow fibers, and the like, utilizing a fine space in the material, Any solvent can be used as long as the solvent can be separated.

  Further, if the separation layer has a property with high affinity to the solvent, the efficiency of penetration and passage of the solvent can be increased, and the separation efficiency can be increased.

  For example, when the solvent is water, the surface of the solvent separation layer is preferably hydrophilic. This hydrophilic solvent separation layer can be formed by using a porous material made of a hydrophilic material or by performing a surface treatment on the porous material. For example, when the porous material is an inorganic material, surface treatment (hydrophilization) with a silane coupling agent or the like is effective.

  In the present invention, the liquid sample is introduced into the flow path and contacts the solvent separation layer with pressure. By utilizing the pressure of the liquid sample, it is possible to more effectively perform separation using the solvent separation layer. Therefore, the side surface of the flow channel in the present invention means a wall surface in the flow channel, and includes all of the upper surface, the lower surface, the lateral surface, and the like.

(Solvent retention layer)
The solvent holding layer is installed via the solvent separation layer. The solvent holding layer in the present invention means a layer having a function of holding the solvent separated from the liquid sample by the solvent separation layer.

  Therefore, if a space for holding the solvent is formed, a solvent holding layer can be formed, but it is more preferable that a material having a high solvent absorption capacity is provided. For example, when the solvent is water, a water-absorbing polymer, a water-absorbing gel, a porous material, a nonwoven fabric, or the like can be used as a material having a high solvent absorption capacity.

  Although these materials can obtain an effect as long as they are provided in at least a part of the solvent holding layer, it is more desirable that the material is in contact with the solvent separation layer and directly absorbs the solvent that has passed through the solvent separation layer. .

  In order to increase the solvent introduction efficiency, it is desirable that the solvent retention layer has air holes that allow air existing inside the solvent retention layer to escape. And in order to raise efficiency further, you may provide the means to make a solvent retention layer into a pressure reduction state.

  Further, a part of the solvent holding layer may be a solvent separation layer, and one member may serve as both layers.

  For example, by using a water-absorbing gel that selectively absorbs water or a component having a molecular weight equal to or less than the analytical component, it is possible to have both a function of separating the solvent from the liquid sample and a function of retaining the separated solvent. . Therefore, the water-absorbing gel layer serves as both the solvent separation layer and the solvent holding layer.

(Arrangement of flow path, solvent separation layer, solvent retention layer)
The concentration element according to the present invention includes a flow path, a solvent separation layer, and a solvent holding layer, and the solvent separation layer is disposed in contact with at least a part of a side surface of the flow path, and the solvent holding layer is interposed via the solvent separation layer. Installed.

  Hereinafter, these arrangements will be described in detail with reference to the drawings.

  4, 5, and 6 are schematic cross-sectional views showing examples of arrangement of the flow path (3), the solvent separation layer (23, 33), and the solvent holding layer (24, 34).

  In FIG. 4, a solvent holding layer (24) is provided on a first substrate (21), the solvent holding layer is partially removed to form a groove (14), and a solvent separation layer (23 ).

  The groove portion (14) is coated with a material that becomes a solvent separation layer to form a solvent separation layer (23), or the groove portion is filled with a material that becomes a solvent separation layer, and one of the formed solvent separation layers (23) is formed. Remove the part. Thereafter, the channel (3) can be formed by superimposing the second substrate (22).

  This arrangement is effective when, for example, the solvent holding layer (24) is filled with a solvent absorbent having a relatively high strength and excellent workability (removability).

  In FIG. 5, a solvent holding layer (24) is provided on the first substrate (21) in the same manner as in FIG. 4, and the groove (14) is formed by partially removing the solvent holding layer. FIG. 6 is a schematic view showing an example in which a solvent holding layer is installed as a separate member (16) on the substrate (21) and a gap is formed as a groove (14).

  The solvent separation layer (23) is provided on the surface of the groove by an operation such as coating of a material to be the solvent separation layer or removal after filling.

  FIG. 6 is a schematic view showing an example in which a solvent holding layer (34) is provided on the first substrate (31) and the flow path (3) is formed by the other member (16).

  In FIG. 6 (a), a solvent holding layer (34) and a solvent separation layer (33) are provided on a first substrate (31), and a second substrate (32) provided with a groove (14) is provided. It is possible to form the flow path (3) by overlapping.

  FIG. 6 (b) shows a state in which another member (16) is sandwiched between a first base (31) and a second base (32) each having a solvent holding layer (34) and a solvent separation layer (33). Together, it is possible to form the flow path (3).

  In FIG. 6 (c), a solvent holding layer (34) and a solvent separation layer (33) are further provided on both the first substrate (31) and the second substrate (32), and another member (16) is sandwiched between them. In this example, the flow paths (3) are formed by superimposing them.

  These arrangements are simple in terms of the production method because it is not necessary to perform fine processing on the solvent holding layer (34) and the solvent separation layer (33). In addition, since the solvent separation layer and the solvent holding layer are arranged on the lower surface in the flow path, it is possible to increase the solvent separation efficiency when the liquid sample is introduced into the flow path and concentrated.

  In addition, you may use combining the structure typically shown in FIG.4, FIG.5, FIG.6. These configurations are examples, and the present invention is not limited to these configurations as long as the effects of the present invention can be obtained.

(Chemical analyzer)
The chemical analysis apparatus according to the present invention is a chemical analysis apparatus that performs a chemical analysis using a liquid sample, and includes the concentration element and means for introducing the liquid sample into the concentration element. The liquid sample is introduced into the concentration element and concentrated by the means for introducing the liquid sample.

  The chemical analysis apparatus according to the present invention mainly refers to a chemical apparatus integrated and miniaturized on a chip, called so-called μTAS, lab-on-chip, or microreactor. Therefore, it includes not only an analysis but also a device for performing a chemical reaction.

  The concentration element has a simple configuration as described above, and can be easily formed on a substrate. Therefore, it is suitable for the chemical analyzer according to the present invention.

  7 and 8 are schematic views showing examples of the analyzer according to the present invention.

  7 includes a liquid sample injection tank (40), a liquid feed channel (41), an on-off valve (42, 51), a pump (43, 52), a connection channel (on a substrate (10)). 44), a concentration element (50), a detection part (53), and a liquid sample discharge tank (54).

  For example, the liquid sample is introduced from the liquid sample injection tank (40), passes through the liquid supply channel (41) and the connection channel (44), and is introduced into the concentration element (50). In the present invention, as described above, the concentration element is formed on the substrate, and the connection channel is formed as the liquid sample introduction means, whereby the liquid sample can be introduced and concentrated.

  Furthermore, it is desirable that a fluid control element such as a pump or an on-off valve is provided. By changing the flow rate of the liquid sample, the pressure and the residence time of the liquid in the concentration element can be controlled, and the concentration rate can be changed. It is also possible to enhance the concentration effect by closing the on-off valve 2 (51) and applying pressure in the concentration element.

  A plurality of these concentration elements and introduction means may be formed on the substrate. It is also possible to enhance the concentration effect by using a plurality of concentration elements for one type of liquid sample. Further, a plurality of concentration elements may be formed for a plurality of liquid samples, and concentration may be performed for each.

  When forming a plurality of concentration elements, a solvent flow path, a solvent separation layer, and a solvent holding layer may be formed separately for each of the concentration elements. The solvent holding layer may also be used.

  In the chemical analyzer according to the present invention, it is further preferable that a detection unit is provided on the substrate. The concentrated liquid sample is introduced into the detection unit and can be analyzed.

  In the detection unit, a chemical or physical change due to the presence of the analysis component is detected. For example, an analysis can be performed by detecting changes in current, voltage, light quantity, mass, heat quantity, and the like.

  The detecting means for detecting these changes may be provided on the substrate, or may be provided outside. For example, it is desirable that an electrode or the like for detecting an electrical change is provided on the base, but a light receiving means or the like for detecting an optical change may not be provided on the base.

  In addition, these changes may occur when the concentrated analysis component causes some chemical reaction in the detection unit.

  For example, a change caused by a reaction between an analysis component and another substance provided in the detection unit may be detected in advance, or a change that occurs as a result of another substance being introduced into the detection unit and reacting with the analysis component may be detected. . Further, a change may occur as a result of reaction of the analysis component with light, heat, or the like applied to the reaction part.

  Note that the concentration element may also serve as the detection unit. For example, it is also possible to perform detection at the same time using a chemical analysis apparatus configured as shown in FIG. 8 while introducing a liquid sample into the concentration element and removing the solvent.

  The chemical analysis apparatus according to the present invention can be suitably used as a so-called biosensor.

  A biosensor is a measurement device that utilizes the excellent molecular recognition ability of living organisms and biomolecules. Therefore, it is desirable that the detection unit is provided with a substance (capturing component) having excellent molecular recognition ability.

  The capture component is immobilized on the detection unit, the capture component recognizes the analysis component, and detects chemical and physical changes that occur when the capture component is captured, whereby the chemical analysis apparatus according to the present invention is used as a biosensor. It can be used.

  The trap component may be any substance that is related to the selection of the analysis component in the liquid sample.For example, a substance that selectively reacts directly with the analysis component in the liquid sample (so-called receptor), a substance that is related to the reaction of the analysis component ( For example, a substance that selectively catalyzes the reaction of the analysis component), a substance that inactivates a substance other than the analysis component in the liquid sample, and the like.

  The capturing body component may also have a function related to the presence / absence and degree of detection, for example, a function of reacting with a substance released by a receptor or a remaining substance to develop a color. Examples of the capture component used in the present invention include, but are not limited to, enzymes, sugar chains, catalysts, antibodies, antigens, genes, color reagents, and the like.

  Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples, and the functions intended by the present invention, such as materials, composition conditions, reaction conditions, and the like. The concentration element and the chemical analysis apparatus can be freely changed within a range in which the concentration element and the chemical analysis apparatus can be obtained.

  Hereinafter, Examples 1, 2, 3, and 4 show examples in which a concentration element is prepared and a protein aqueous solution is concentrated, and Example 5 shows an example in which a chemical analyzer is prepared and analyzed.

(Example 1)
This example is an example in which a concentration element having a channel (3) having a shape as shown in FIG. 9 is produced and the concentration process is performed. 9A is a plan view taken along the line AA ′ (viewed from above the base), and FIG. 9B is a longitudinal sectional view taken along the line BB ′.

  Even in a concentration element having a very simple structure as in this embodiment, it is possible to obtain a concentration effect.

  FIG. 10 is a process diagram showing a production process of the concentration element of this example.

  First, a silicon substrate is prepared and used as a first substrate (31) (FIG. 10 (a)).

  Next, a solvent holding layer (34) is formed on the first substrate. In this example, a polyolefin-based hydrophilic porous sintered compact Sunfine (manufactured by Asahi Kasei Co., Ltd.) is used as the water-absorbing material, and is bonded to the first substrate (31) (FIG. 10 (b)).

  Next, as the solvent separation layer (33), a cellulose semipermeable membrane is placed on the solvent retention layer (34) (FIG. 10 (c)).

  Further, a resin substrate having a groove (14) is prepared as the second base (32), and the flow path (3) is formed by pressure bonding (FIG. 10 (d)).

  In order to prevent liquid leakage and increase the strength of the element, the periphery is sealed with a resin material. However, the inlet (2) and the outlet (4) are not sealed, and a part of the solvent holding layer (34) is left as an open part so as to be an air hole.

  A connection flow path (not shown) is formed at the introduction port (2) and the discharge port (4), and a silicon tube is connected, and the connection portion is sealed with a resin material so that there is no liquid leakage. The silicon tube on the inlet (2) side is connected to a disposable syringe, and the flow rate can be controlled by using a syringe pump.

  Through the above operation, the concentration element as shown in FIG. 9 can be produced. The flow path of the concentrating element in this example has a width of 300 μm and a height of 50 μm.

  Next, an example of concentrating a liquid sample using this concentration element will be described.

  First, a PBS solution of 10 μmol / L anti-BSA antibody is prepared (PBS buffer: 0.1 mol / L NaCl, 10 mmol / L sodium phosphate (pH 7.4)).

  This solution is introduced into the concentration element from the connection channel and the introduction port using a syringe. By using one surface of the fine channel as a solvent separation layer as in this example, the ratio of the area in contact with the solvent separation layer to the volume of the liquid sample is increased, and the concentration effect can be enhanced. .

  In addition, by changing the flow rate of the liquid sample using a syringe pump, the pressure and the residence time of the liquid in the concentration element can be controlled, and the concentration rate can be changed by these controls. .

  The concentration rate can be determined by measuring the absorbance of the solution discharged from the outlet and determining the concentration of the anti-BSA antibody. In the present embodiment, it is possible to obtain a concentration ratio of 2 times or more.

(Example 2)
In this example, a concentration element having a flow path (3) having a shape as shown in FIG. 11 was produced and the concentration process was performed.

  11A is a plan view taken along the line AA ′ (viewed from above the base), and FIG. 11B is a longitudinal sectional view taken along the line BB ′. The present embodiment is an example of a concentration element in which the flow path (3) includes an introduction port (2), and the cross-sectional area of the introduction port (2) is smaller than the cross-sectional area of the flow path (3).

  A manufacturing method is described.

  First, a silicon substrate is prepared and used as the first substrate (31). Next, a solvent holding layer (34) is formed on the first substrate. Next, as the solvent separation layer (33), a cellulose semipermeable membrane is placed on the solvent holding layer (34). The above operations are the same as those in the first embodiment.

  Further, a resin substrate having a groove is prepared as the second base body (32), and the flow path (3) is formed by pressure bonding. However, the second base body (32) in this example has an additional space for the introduction port (2), the discharge port (4), and the connection flow channel (5) in addition to the groove portion serving as the flow channel (3). Is formed.

  In order to prevent liquid leakage and increase the strength of the element, the periphery is sealed with a resin material. However, the inlet (2) and the outlet (4) are not sealed, and a part of the solvent holding layer (34) is left as an open part so as to be an air hole.

  Further, a silicon tube is connected to the connection channel (5), and the connection portion is sealed with a resin material so that there is no liquid leakage. The silicon tube on the inlet side is connected to a disposable syringe, and the flow rate can be controlled by using a syringe pump.

  Through the above operation, a concentration element as shown in FIG. 11 can be produced. In this embodiment, the flow path of the concentration element is 500 μm wide and 100 μm high, and the inner diameter of the inlet is 70 μm.

  Next, an example of concentrating a liquid sample using this concentration element will be described.

  As in Example 1, a PBS solution of anti-BSA antibody is introduced into the concentration element. In the present invention, the cross-sectional area of the concentration element channel (3) is larger than the cross-sectional area of the introduction port (2), and it is possible to take a longer time for the introduced solution to stay in the concentration element.

  Thus, for example, when the concentration element (50) is formed on the same substrate (10) as the other elements, the liquid sample (1) is flowed at a constant flow rate, and the different elements are sequentially operated in a plurality of elements, the concentration element ( It is possible to increase the concentration effect by increasing the residence time in 50).

  The concentration rate can be determined by measuring the absorbance of the solution discharged from the outlet and determining the concentration of the anti-BSA antibody. In this embodiment, it is possible to obtain a concentration ratio of 3 times or more.

Example 3
In this example, a concentration element having a flow path (3) having a shape as shown in FIG. 12 was produced and the concentration process was performed.

  12A is a plan view taken along the line AA ′ (viewed from above the base), and FIG. 12B is a longitudinal sectional view taken along the line BB ′. This embodiment is an example of a concentration element in which the flow path (3) includes an inlet (2) and an outlet (4), and the sectional area of the outlet (4) is smaller than the sectional area of the inlet (2). .

  A manufacturing method is described.

  First, a silicon substrate is prepared and used as the first substrate (31). Next, a solvent holding layer (34) is formed on the first substrate. Next, as the solvent separation layer (33), a cellulose semipermeable membrane is placed on the solvent holding layer (34). The above operations are the same as those in the first embodiment.

  Further, a resin substrate having a groove is prepared as the second base body (32), and the flow path (3) is formed by pressure bonding. However, the second substrate in the present embodiment is formed such that the cross-sectional area of the discharge port (4) is smaller than the cross-sectional area of the introduction port (2), the shape of the groove portion serving as the flow path (3). Yes.

  In order to prevent liquid leakage and increase the strength of the concentration element, the periphery of the concentration element is sealed with a resin material. However, the inlet (2) and the outlet (4) are not sealed, and a part of the solvent holding layer (34) is left as an open part so as to be an air hole.

  A connection flow path (not shown) is formed at the introduction port (2) and the discharge port (4). Further, a silicon tube is connected, and the connection portion is sealed with a resin material so as not to leak. The silicon tube on the inlet (2) side is connected to a disposable syringe, and the flow rate can be controlled by using a syringe pump.

  Through the above operation, the concentration element as shown in FIG. 12 can be produced. The flow path of the concentrating element in this embodiment is 300 μm wide at the inlet (2), 100 μm wide at the outlet, and 50 μm high.

  Next, an example of concentrating a liquid sample using this concentration element will be described.

  As in Example 1, a PBS solution of anti-BSA antibody is introduced into the concentration element. In the present invention, the cross-sectional area of the discharge port (4) is smaller than the cross-sectional area of the introduction port (2) of the flow path. Therefore, even if the solvent is removed in the concentration element and the amount of liquid decreases, it is possible to reduce the change in flow rate.

  The concentration rate can be determined by measuring the absorbance of the solution discharged from the discharge port (4) and determining the concentration of the anti-BSA antibody. In this embodiment, it is possible to obtain a concentration ratio of 3 times or more.

Example 4
In this example, a concentration element having a flow path (3) having a shape as shown in FIG. 13 was produced and the concentration process was performed.

  FIG. 13A is a plan view taken along the line AA ′ (viewed from above the base), and FIG. 13B is a longitudinal sectional view taken along the line BB ′. The present embodiment is an example of a concentration element in which the flow path (3) includes a throttle as the pressure loss part (6).

  A manufacturing method is described.

  First, a silicon substrate is prepared and used as the first substrate (31). Next, a solvent holding layer (34) is formed on the first substrate. Next, as the solvent separation layer (33), a cellulose semipermeable membrane is placed on the solvent holding layer (34). The above operations are the same as those in the first embodiment.

  Further, a resin substrate having a groove is prepared as the second base body (32), and the flow path (3) is formed by pressure bonding. However, the second substrate in the present embodiment is formed by providing a projection serving as a restriction in a groove serving as the flow path (3).

  In order to prevent liquid leakage and increase the strength of the concentration element, the periphery of the concentration element is sealed with a resin material. However, the inlet (2) and the outlet (4) are not sealed, and a part of the solvent holding layer (34) is left as an open part so as to be an air hole.

  A connection flow path (not shown) is formed at the introduction port (2) and the discharge port (4). Further, a silicon tube is connected, and the connection portion is sealed with a resin material so as not to leak. The silicon tube on the inlet side is connected to a disposable syringe, and the flow rate can be controlled by using a syringe pump.

  Through the above operation, the concentration element as shown in FIG. 13 can be manufactured. In the present embodiment, the flow path of the concentration element has a width of 400 μm and a height of 50 μm, and the width is narrowed to 70 μm by installing a diaphragm.

  Next, an example of concentrating a liquid sample using this concentration element will be described.

  As in Example 1, a PBS solution of anti-BSA antibody is introduced into the concentration element. In the present invention, since the restrictor is provided in the flow path (3), the restrictor becomes a resistance obstacle to the flow of the liquid, and the concentration effect can be increased.

  The concentration rate can be determined by measuring the absorbance of the solution discharged from the outlet and determining the concentration of the anti-BSA antibody. In this embodiment, it is possible to obtain a concentration ratio of 4 times or more.

(Example 5)
This example is an example in which a chemical analyzer having the configuration shown in FIG. 7 is produced, and an aqueous protein solution is concentrated to detect the protein.

  A glass substrate is used for the substrate (10), and the method for producing the concentration element (50) is in accordance with Example 4. In the detection unit (53), BSA molecules are immobilized in advance as a capturing component. This immobilization process can be performed by adding a BSA molecule after the amination process is performed on the detection unit.

  Specifically, an ethanol solution of aminopropyltrimethoxysilane (manufactured by Aldrich)) is dropped on a glass substrate serving as the detection unit (53) and allowed to stand, and then washed with ethanol and dried. By this process, an amino group is introduced into the detection unit.

  Further, a 100 μmol / L BSA aqueous solution is dropped onto the detection unit, dried and fixed, and then immersed in a 5% skim milk solution for 1 hour and dried at room temperature. Through the above processing, the BSA molecule is fixed to the detection unit.

  If it is desired to further prevent the BSA molecule from being immobilized to other than the detection unit (53), the above process may be performed by masking the region other than the detection unit.

  In contrast, as a comparative example, a chemical analyzer without a concentrating element is produced at the same time.

  Next, an example of protein detection using this chemical analyzer and a chemical analyzer produced as a comparative example will be described.

  First, a 100 μmol / L Cy3-labeled anti-BSA antibody PBS solution is introduced into the chemical analyzer according to this example. Then, PBS buffer is introduced and washed. The presence of the anti-BSA antibody can be detected by observing the detection part with a fluorescence microscope and observing the fluorescence.

  By using the chemical analysis apparatus according to the present embodiment, it is possible to observe strong fluorescence compared to the comparative example. Furthermore, the time required for observing fluorescence with sufficient intensity for observation can also be shortened.

  As described above, according to the present invention, it is possible to detect an analysis component in a short time and with high sensitivity.

It is a schematic diagram which shows the example of the shape of the flow path with which the concentration element which concerns on this invention is equipped. It is a schematic diagram which shows the example of the flow-path cross section by this invention. It is a schematic diagram which shows the example of the formation method of a groove part. It is the cross-sectional schematic diagram which showed the example of arrangement | positioning of a flow path, a solvent separation layer, and a solvent holding layer. It is the cross-sectional schematic diagram which showed the example of arrangement | positioning of a flow path, a solvent separation layer, and a solvent holding layer. It is the cross-sectional schematic diagram which showed the example of arrangement | positioning of a flow path, a solvent separation layer, and a solvent holding layer. It is the schematic which shows the structural example of the chemical analyzer by this invention. It is the schematic which shows the structural example of the chemical analyzer by this invention. 1 is a schematic diagram showing a concentration element according to Example 1. FIG. FIG. 6 is a process diagram showing a production process of the concentration element according to Example 1. 6 is a schematic diagram showing a concentrating element according to Example 2. FIG. 6 is a schematic diagram showing a concentrating element according to Example 3. FIG. 6 is a schematic diagram showing a concentrating element according to Example 4. FIG.

Explanation of symbols

1: Liquid sample 2: Inlet port 3: Channel 4: Discharge port
10: Base
21, 31: First substrate
22, 32: Second substrate
23, 33: Solvent separation layer
24, 34: Solvent retention layer

Claims (8)

A concentration element for concentrating a liquid sample,
A flow path formed on the substrate;
A concentration element comprising: a solvent separation layer disposed in contact with at least a part of a side surface of the flow path; and a solvent holding layer disposed via the solvent separation layer.   The concentration element according to claim 1, wherein the solvent separation layer is made of a porous material.   The concentration element according to claim 1, wherein the solvent holding layer includes a material that absorbs the solvent.   The concentration element according to claim 1, wherein the flow path includes an inlet for introducing a liquid sample, and the cross-sectional area of the inlet is smaller than the cross-sectional area of the flow path.   5. The flow path according to claim 1, wherein the flow path includes an inlet for introducing a liquid sample and an outlet for discharging the liquid sample, and a cross-sectional area of the outlet is smaller than a cross-sectional area of the inlet. The concentration element according to any one of the above.   The concentration element according to claim 1, wherein the flow path includes a pressure loss portion. A chemical analyzer for performing chemical analysis using a liquid sample,
A concentration element comprising: a channel formed on a substrate; a solvent separation layer installed in contact with at least a part of a side surface of the channel; and a solvent holding layer installed via the solvent separation layer;
A chemical analysis apparatus comprising: an introduction unit that introduces the liquid sample into the concentration element.   The chemical analysis apparatus according to claim 7, further comprising a detection unit that detects the presence or absence or amount of a substance in the liquid sample.

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