JP6925926B2 - Microchips, analytical systems and methods for manufacturing microchips - Google Patents

Description

The present invention relates to a microchip, an analysis system and a method for manufacturing a microchip.

Specimen analysis by capillary electrophoresis has been performed conventionally, and in recent years, electrophoresis using a microchip made into a chip device has been performed in order to reduce the size and simplification of the apparatus. Further, as the microchip, from the viewpoint of improving the separation speed and the separation accuracy of the sample, it is preferable to fix the cationic polymer on the inner wall of the analysis part such as the capillary tube and introduce the positive charge layer.

For example, a cationic polymer having a quaternary onium group is immobilized on at least one surface of each of the pair of resin substrates, and the resin substrate is bonded using the surface on which the cationic polymer is immobilized as a bonding surface. A method for manufacturing a chip provided with a microchannel including the above has been proposed (see, for example, Patent Document 1).

Further, as an analysis device used for sample analysis, a first unit having a sampling unit and a second unit having an analysis unit such as a capillary tube may be separately provided, and the user may use the first unit. After collecting a sample sample (sample) using one unit, the first unit is set in the second unit and sample analysis is performed.

In general, since the sampling section sucks the sample by capillary force, it is desirable that the inner wall of the sampling section has high hydrophilicity. Therefore, it is preferable to hydrophilize the inner wall of the sampling portion. For example, a hydrophilic film in which a composition containing a hydrolyzable organic silicon compound and inorganic compound fine particles is cured is formed on the surface of the resin molded product to form a resin molded product. A method of hydrolyzing the surface has been proposed (see, for example, Patent Document 2).

Japanese Unexamined Patent Publication No. 2015-166127 Japanese Unexamined Patent Publication No. 2005-126460

In an analysis device (microchip) having a first unit having a sampling unit and a second unit having an analysis unit such as a capillary tube separately, a separate unit has a sampling unit and an analysis unit. Therefore, it is necessary to manufacture each of the two units, which increases the production cost. Therefore, from the viewpoint of suppressing production costs, it is preferable that the same unit includes a sampling unit and an analysis unit.

Here, as described above, since the sampling unit sucks the sample by the capillary force, it is preferable that the sampling unit has high hydrophilicity, and the analysis unit has analysis performance (separation) especially when it is a microchannel such as a capillary tube. It is preferable to have a positive charge layer in order to improve the speed, separation accuracy, etc.). From the viewpoint of suppressing the production cost of the chip device, a method for producing a microchip capable of forming a sampling unit to which hydrophilicity is imparted and an analysis unit to which a positive charge layer is provided in the same process is desirable.

However, although the method described in Patent Document 1 can stably impart a positive charge layer to a microchannel or the like, it cannot impart high hydrophilicity. Further, in the method described in Patent Document 2, although it is possible to impart high hydrophilicity to the microchannels and the like, it is not possible to impart a stable positive charge layer.

The present invention includes a microchip provided with a sampling unit and an analysis unit, and the sampling unit and the analysis unit are provided with both a hydrophilic layer and a positive charge layer, an analysis system provided with the above-mentioned microchip, and the above-mentioned analysis system. It is an object of the present invention to provide a method for manufacturing a microchip.

Specific means for achieving the above-mentioned objectives are as follows.
<1> Immobilization of a cationic polymer in which a hydrophilic substance is bound to a region where a sampling part for collecting a sample is formed and a region where an analysis part for analyzing the sample is formed on a pair of base materials. The sampling unit, which comprises a step and a joining step of joining the pair of base materials after the immobilization step, wherein the cationic polymer to which the hydrophilic substance is bound is immobilized by the joining step. And a method for manufacturing a microchip on which the analysis unit is formed.
<2> The immobilization step includes a modification step of immobilizing the cationic polymer in a region where the sampling unit is formed and a region where the analysis unit is formed, and the immobilization step of the cationic polymer. The method for producing a microchip according to <1>, which comprises a hydrophilization step of binding a hydrophilization substance.
<3> In the immobilization step, after the hydrophilic substance was bound to the cationic polymer, the hydrophilized substance was bound to the region where the sampling portion was formed and the region where the analysis portion was formed. The method for producing a microchip according to <1>, wherein the cationic polymer is immobilized.
<4> The method for producing a microchip according to any one of <1> to <3>, wherein the hydrophilic substance has one or more anionic functional groups.
<5> The method for producing a microchip according to <4>, wherein the anionic functional group contains at least one of a carboxy group and a sulfo group.
<6> The method for producing a microchip according to any one of <1> to <5>, wherein the hydrophilic substance has one or more hydrophilic functional groups.
<7> The method for producing a microchip according to <6>, wherein the hydrophilic functional group contains at least one of a hydroxyl group and an amino group.
<8> The method for producing a microchip according to any one of <1> to <7>, wherein the cationic polymer contains at least one of polyquaternium and a dimethylamine-epichlorohydrin copolymer.

<9> A sampling unit for collecting a sample and an analysis unit for analyzing the sample are provided, and a cationic polymer in which a hydrophilic substance is bound is immobilized on the sampling unit and the inner wall of the analysis unit. Microchip.
<10> The microchip according to <9>, wherein one unit includes the sampling unit and the analysis unit.
<11> The microchip according to <9> or <10>, wherein the sampling unit and the analysis unit are sites formed by joining a pair of base materials.
<12> Of <9> to <11>, the cationic polymer immobilized on the inner wall of the sampling unit and the cationic polymer immobilized on the inner wall of the analysis unit are the same. The microchip described in any one.
<13> The microchip according to any one of <9> to <12>, wherein the hydrophilic substance has one or more anionic functional groups.
<14> The microchip according to <13>, wherein the anionic functional group contains at least one of a carboxy group and a sulfo group.
<15> The microchip according to any one of <9> to <14>, wherein the hydrophilic substance has one or more hydrophilic functional groups.
<16> The microchip according to <15>, wherein the hydrophilic functional group contains at least one of a hydroxyl group and an amino group.
<17> The microchip according to any one of <9> to <16>, wherein the cationic polymer contains at least one of polyquaternium and a dimethylamine-epichlorohydrin copolymer.
<18> The microchip according to any one of <9> to <17>, wherein the sampling unit collects the sample using capillary force.
<19> The microchip according to any one of <9> to <18>, which is a capillary tube.
<20> The microchip according to any one of <9> to <19> used for analysis of a biological substance in the sample by electrophoresis.
<21> The microchip according to <20>, wherein the biological substance is hemoglobin.

<22> An analysis system including the microchip according to any one of <9> to <21> and an analyzer to which the microchip is attached.

According to one aspect of the present invention, an analysis system including a sampling unit and an analysis unit, a microchip having both a hydrophilic and positively charged layer added to the sampling unit and the analysis unit, and the above-mentioned microchip. , And the above-mentioned method for manufacturing a microchip can be provided.

It is a schematic block diagram of the base material (chip part A) used in the method of manufacturing a microchip of one aspect of this invention, (a) is a top view, (b) is a bottom view (a view from a joint surface). (C) is a cross-sectional view taken along the line A'-A'of (a). It is the schematic block diagram of the base material (chip part B) used in the manufacturing method of the microchip of one aspect of this invention, (a) is the top view, (b) is the bottom view (the view from the joint surface). (C) is a cross-sectional view taken along the line B'-B'of (a). It is a schematic block diagram of the microchip manufactured by the method of manufacturing the microchip of one aspect of this invention, (a) is a top view, (b) is a C'-C'line sectional view of (a). .. It is a graph which shows the result of having analyzed the hemoglobin in the whole human blood using the microchip manufactured in each Example and each comparative example.

Hereinafter, a method for manufacturing a microchip according to one aspect of the present invention and the microchip will be described.
In the present specification, the numerical range represented by using "~" means a range including the numerical values before and after "~" as the lower limit value and the upper limit value.

[Manufacturing method of microchip]
In the method for producing a microchip according to one aspect of the present invention, a hydrophilic substance is bound to a region in which a sampling portion for collecting a sample is formed and a region in which an analysis portion for analyzing the sample is formed on a pair of substrates. The cationic polymer to which the hydrophilized substance is bound by the joining step includes an immobilization step of immobilizing the cationic polymer and a joining step of joining the pair of base materials after the immobilization step. The sampling unit and the analysis unit are formed.
It is possible to impart both hydrophilic and positively charged layers to the region where the sampling part is formed and the region where the analysis part is formed in the pair of substrates in the immobilization step, and joining after the immobilization step. In the process, a pair of base materials can be joined to form a sampling section and an analysis section in which a cationic polymer to which a hydrophilic substance is bound is immobilized. Further, in a series of steps, it is possible to impart the properties required for each of the sampling unit and the analysis unit to the region to be the sampling unit and the region to be the analysis unit in the immobilization step. The process is simplified, and the production cost of the microchip can be reduced.

The microchip obtained by the method for producing a microchip of this embodiment and the microchip of this embodiment described later are used in various analytical methods for various samples, and are preferably electrophoresis (preferably capillary electrophoresis). ) Is used for the analysis of substances (preferably biological substances) in the sample. Examples of the sample include samples of biological blood, urine, sweat, etc., and samples subject to environmental surveys such as water quality surveys and geological surveys. The sample is preferably blood collected from the human body or the like. Further, as the analysis target, a biological substance contained in the sample is preferable, and a biological substance contained in blood is more preferable. Hemoglobin is preferable among biological substances.

Among the components contained in blood, the components to be analyzed include hemoglobin (Hb), albumin (Alb), gamma globulin (α1, α2, β, gamma globulin), fibrinogen and the like. Examples of hemoglobin include normal hemoglobin (HbA0), glycated hemoglobin, modified hemoglobin, fetal hemoglobin (HbF)), mutant hemoglobin, and the like. Examples of the saccharified hemoglobin include hemoglobin A1a (HbA1a), hemoglobin A1b (HbA1b), hemoglobin A1c (HbA1c), GHbLys and the like. Examples of the hemoglobin A1c include stable HbA1c (S-HbA1c) and unstable HbA1c (L-HbA1c). Examples of the modified hemoglobin include carbamylated Hb and acetylated Hb.

<Immobilization process>
In the immobilization step, the cationic polymer in which the hydrophilic substance is bound is immobilized in the region where the sampling portion is formed and the region where the analysis portion is formed on the pair of substrates. In this step, the cationic polymer in which the hydrophilic substance is bound is immobilized in the region where the sampling part is formed and the region where the analysis part is formed, and in the next joining step, the sampling part and the analysis part are subjected to. A microchip with both hydrophilic and positively charged layers is formed.

(Base material)
As the base material, for example, a pair of base materials on which a sampling unit for collecting a sample by joining and an analysis unit for analyzing the sample are formed is used. Examples of the pair of base materials include chip component A and chip component B, which are substantially elongated rectangular plate-shaped members, as shown in FIGS. 1 and 2.

As shown in FIG. 1, the chip component A includes through holes 11 to 14, a detection window 15, and a first groove 16 (a region in which the sampling unit 1 is formed). Further, as shown in FIG. 2, the chip component B includes a detection window 15 and a second groove 18 (a region in which the analysis unit 2 is formed). Further, the chip component A has a region 19 in which the analysis unit 2 is formed between the through hole 13 and the through hole 14 on the surface shown in FIG. 1 (b), and the chip component B has a region 19 in which the analysis unit 2 is formed. The surface shown in (b) of No. 1 has a region 17 in which the sampling unit 1 is formed. Then, in the joining step, the surfaces shown in FIGS. 1B and 2B are used as the joining surfaces, the first groove 16 and the region 17 face each other, and the second groove 18 and the region 19 are opposed to each other. By joining the chip component A and the chip component B so as to face each other, a microchip 100 having a sampling unit 1 and an analysis unit 2 is formed as shown in FIG.

The through holes 11 to 14 in the chip component A penetrate the chip component A in the thickness direction, and each of the through holes penetrates the sample collection port 3, the sample arrival port 4, the sample holding portion 5, and the migration in the microchip 100. Corresponds to the liquid holding unit 6.

The detection window 15 in the chip component A and the chip component B is for injecting and emitting light to be used for analysis when the microchip 100 is analyzed by electrophoresis, and is used, for example, for measuring absorbance. .. In the chip component B, the detection window 15 is arranged so as to face at least a part of the second groove 18.

The first groove 16 in the chip component A corresponds to the region where the sampling portion 1 is formed, and is provided between the through hole 11 and the through hole 12 on the surface shown in FIG. 1 (b). It is a groove. In the immobilization step, the cationic polymer to which the hydrophilic substance is bound is immobilized on the wall surface of the first groove 16 to impart a hydrophilic and positive charge layer. Further, in the joining step described later, the first groove 16 in the chip component A and the region 17 in the chip component B are joined so that the chip component A and the chip component B face each other, whereby the first groove is formed. The sampling unit 1 is formed at a position where the 16 and the region 17 face each other.

The first groove 16 is preferably a microchannel provided between the through hole 11 and the through hole 12. The size of the first groove 16 is not particularly limited, and for example, the depth is preferably 100 μm to 1000 μm, the width is 100 μm to 2000 μm, and the length is preferably 1 mm to 20 mm.

The region 19 in the chip component A corresponds to the region in which the analysis unit 2 is formed, and is a region provided between the through hole 13 and the through hole 14 on the surface shown in FIG. 1 (b). In the immobilization step, the cationic polymer to which the hydrophilic substance is bound is immobilized in the region 19 to impart a hydrophilic and positive charge layer. Further, in the joining step described later, the region 19 and the chip component B are joined so that the region 19 in the chip component A and the second groove 18 in the chip component B face each other. The analysis unit 2 is formed at a position facing the groove 18 of 2.

The second groove 18 in the chip component B corresponds to the region where the analysis unit 2 is formed, and when the chip component A and the chip component B are joined, the through holes 13 and the through holes 13 are formed at both ends of the second groove 18. The sample holding part 5 and the running solution holding part 6 corresponding to the holes 14 are provided so as to be located. In the immobilization step, the cationic polymer to which the hydrophilic substance is bound is immobilized on the wall surface of the second groove 18, and a hydrophilic and positive charge layer is imparted.

The second groove 18 functions as an analysis unit that analyzes the sample when the chip component A and the chip component B are joined, preferably as a capillary tube that analyzes the sample by electrophoresis. Further, the second groove 18 extends linearly along the longitudinal direction of the chip component B. The size of the second groove 18 is not particularly limited, and for example, the depth is preferably 25 μm to 100 μm, the width is 25 μm to 100 μm, and the length is 5 mm to 150 mm.

The region 17 in the chip component B corresponds to the region in which the sampling unit 1 is formed, and is a region provided on the surface shown in FIG. 2B. In the immobilization step, the cationic polymer to which the hydrophilic substance is bound is immobilized in the region 17, and a hydrophilic and positively charged layer is imparted. Further, in the joining step described later, the first groove 16 in the chip component A and the region 17 in the chip component B are joined so that the chip component A and the chip component B face each other, whereby the first groove is formed. The sampling unit 1 is formed at a position where the 16 and the region 17 face each other.

Examples of the material of the base material (chip component A and chip component B) include glass, molten silica, resin, etc., from the viewpoint of cost, ease of processing, and ease of immobilization of the cationic polymer. Resin is preferred. Examples of the resin include acrylic resins such as polymethylmethacrylate (PMMA), polymethylmethacrylate, polycarbonate, polyvinylidene chloride, cyclic polyolefin, polyether ether ketone, polystyrene, polytetrafluoroethylene (PTFE), cycloolefin, polypropylene, and Examples thereof include polyethylene, and polymethyl methacrylate is preferable from the viewpoint of excellent light transmission.

(Specific example 1 of the immobilization process)
The immobilization step is a modification step of immobilizing a cationic polymer in a region where a sampling part is formed and a region where an analysis part is formed, and a hydrophilization step in which a hydrophilic substance is bound to the immobilized cationic polymer. The process and may be included. In this case, the cationic polymer is first immobilized in the region where the sampling unit is formed and the region where the analysis portion is formed, and then the hydrophilic substance is bound to the immobilized cationic polymer to form the above region. Immobilizes a cationic polymer to which a hydrophilic substance is bound. As a result, a hydrophilic and positively charged layer can be imparted to the region where the sampling unit is formed and the region where the analysis unit is formed in a series of steps.

For example, a solution 1 containing a cationic polymer is prepared, and the solution 1 is applied to a region in which a sampling portion is formed and a region in which an analysis portion is formed in at least a pair of substrates, and the cationic polymer is applied to the region. Immobilize. After immobilizing the cationic polymer in the above region, the pair of substrates may be washed with water or the like, if necessary. Next, a solution 2 containing a hydrophilic substance is prepared, and the solution 2 is applied to a region in which a sampling portion on which a cationic polymer is immobilized is formed and a region in which an analysis portion is formed in at least a pair of substrates. The hydrophilic substance may be bound to the immobilized cationic polymer.

For example, in the modification step, the cationic polymer is immobilized in the first groove 16 in the chip component A and the region 17 in the chip component B corresponding to the region where the sampling unit 1 is formed. Similarly, in the modification step, the cationic polymer is immobilized in the second groove 18 in the chip component B and the region 19 in the chip component A corresponding to the region where the analysis unit 2 is formed. Then, in the hydrophilization step, the hydrophilic substance is bound to the cationic polymer immobilized in the first groove 16 and the region 17, and the cationic polymer is immobilized in the second groove 18 and the region 19. To bind a hydrophilic substance to.

(Specific example 2 of the immobilization process)
In addition, in the immobilization step, after the hydrophilic substance is bound to the cationic polymer, the cationic polymer to which the hydrophilic substance is bound is fixed in the region where the sampling part is formed and the region where the analysis part is formed. You may make it. In this case, for example, a solution 3 containing a cationic polymer and a hydrophilic substance is prepared, and the hydrophilic substance is bound to the cationic polymer in the solution 3 to form a sampling portion on at least a pair of substrates. By applying the solution 3 to the region and the region where the analysis unit is formed, the cationic polymer in which the hydrophilic substance is bound to the region may be immobilized. Thereby, in the same step, the hydrophilic and positively charged layers can be imparted to the region where the sampling unit is formed and the region where the analysis unit is formed.

For example, the first groove 16 in the chip component A, the region 17 in the chip component B, and the analysis unit 2 correspond to the region where the sampling unit 1 is formed after the hydrophilic substance is bound to the cationic polymer. The cationic polymer is immobilized in the second groove 18 in the chip component B and the region 19 in the chip component A corresponding to the region where the is formed.

In Specific Examples 1 and 2 of the immobilization step, as a method of applying the solutions 1 to 3 to the region where the sampling part is formed and the region where the analysis part is formed, for example, coating, dipping, dropping, spraying and the like are used. Can be mentioned. It is sufficient that the solutions 1 to 3 are applied to the region where the sampling unit is formed and the region where the analysis unit is formed, and the solution is applied to a portion other than the above region on the base material (chip component A and chip component B). 1-3 may be given.

As a pre-step of the immobilization step, a liquid phase surface treatment such as an alkaline solution treatment can be performed from the viewpoint of more preferably immobilizing the cationic polymer in the region where the sampling part is formed and the region where the analysis part is formed. Gas phase surface treatment such as vacuum ultraviolet treatment, plasma treatment, corona discharge treatment, frame treatment, ozone treatment and the like may be performed.

(Cationic polymer)
The cationic polymer used in the method for producing a microchip may be a polymer that is immobilized on the microchip and has a cationic group or a group that can be ionized into the cationic group. The cationic polymer may be a homopolymer of a monomer having a cationic group, a copolymer with another monomer, or a condensed polymer. Examples of the cationic group or the group that can be ionized into the cationic group include a group containing nitrogen, for example, a primary amino group, a secondary amino group, a tertiary amino group, an imino group, an onium group, and a hydrazide. From the viewpoint of improving the analytical performance of the microchip, the onium group is preferable, and the quaternary onium group is preferable. Further, the cationic polymer is preferably water-soluble. The cationic polymer is preferably linear from the viewpoint of improving analytical performance. Further, from the viewpoint of improving analytical performance, the cationic polymer preferably has a total of two or more cationic groups and groups that can be ionized into the cationic groups, and preferably has two or more cationic groups. ..
The cationic polymer may be used alone or in combination of two or more.

Examples of the cationic polymer having a primary amino group to a tertiary amino group include polyallylamine, polyvinylamine, polylysine, polyarginine, polyhistidine, polyornithine, polydiallylamine, and polymethyldiallylamine.
Examples of the cationic polymer having an imino group include polyethyleneimine.

^{The onium group may be a cation formed by coordinating H +} or a cation (R ⁺ , R is an alkyl group, etc.) to a compound containing an element having an unshared electron pair, and is an ammonium ion, a phosphonium ion, or the like. Examples thereof include oxonium ion, sulfonium ion, fluoronium ion and chloronium ion. Among them, ammonium ion and phosphonium ion, which are quaternary onium groups, are preferable, and ammonium ion is more preferable. The cationic polymer is preferably a polycationic polymer having two or more onium groups (preferably quaternary onium groups, more preferably ammonium ions) in one molecule of the polymer. Moreover, the cationic polymer may be a salt such as chloride and sulfide.

The cationic polymer preferably contains a polymer having ammonium ions, more preferably contains at least one of a polyquaternium and a dimethylamine-epichlorohydrin copolymer, and more preferably at least one of a polyquaternium and a dimethylamine-epichlorohydrin copolymer. Is more preferable.

In the present disclosure, "polyquaternium" refers to a cationic polymer containing a structural unit derived from a monomer having ammonium ions. Polyquaternium can be found in the INCI (International Nomenclature for Cosmetic Ingredients) directory. Examples of polyquaternium include polyquaternium-6 (poly (diallyldimethylammonium chloride), polyquaternium-7 (copolymer of amine and diallyldimethylammonium chloride), polyquaternium-4 (Diallyldimethylammonium chloride-hydroxyethyl cellulose copolymer), and polyquaternium-22 (copolymer of acrylic acid and diallyldimethylammonium chloride). Etc., Polyquaternium dimethylammonium salt, Polyquaternium-1 (Ethanol, 2,2', 2''-nitrilotris-, polymer with 1,4-dichloro-2-butene and N, N, N', N'-tetramethyl- 2-butene-1,4-diamine), polyquaternium-11 (Copolymer of vinylpyrrolidone and quaternized dimethylaminoethylryl) and polyquaternium-2 (poly [bis (2-chloroethyl) ether-alt-1,3-bis [3- (dimethylamino)) ) propyl] urea]) and the like.

Examples of the cationic polymer having a hydrazide group include aminopolyacrylamide and the like.

The weight average molecular weight of the cationic polymer is preferably 10,000 or more, preferably 50,000 or more, and more preferably 100,000 or more from the viewpoint of improving analytical performance. The weight average molecular weight of the cationic polymer is preferably 500,000 or less, and more preferably 300,000 or less, from the viewpoint of suppressing an increase in the viscosity of the solution.

As the hydrophilic substance used in the method for producing a microchip, a region capable of binding to the above-mentioned cationic polymer, a region in which a sampling portion is formed and a region in which an analysis portion is formed (preferably, a sampling portion is formed). Any substance that can impart hydrophilicity to the region) may be used.

(Hydrophilic substance)
The hydrophilic substance preferably has one or more anionic functional groups from the viewpoint that it can be suitably bonded to a cationic polymer. Examples of the anionic functional group include a carboxy group (carboxylic acid group), a sulfo group (sulfonic acid group), a sulfuric acid ester group, a phosphoric acid ester group, a phosphonic acid group, a silanol group and the like, and a carboxy group and a sulfo group are preferable. .. The anionic functional group may be a salt.

The hydrophilic substance preferably has one or more (preferably one or two) hydrophilic functional groups, and one or more anionic functional groups, from the viewpoint that hydrophilicity can be preferably imparted. It is more preferable to have a group and one or more hydrophilic functional groups, and it is further preferable to have one anionic functional group and one or more hydrophilic functional groups. Examples of the hydrophilic functional group include a hydroxyl group and an amino group, and for example, at least one of a hydroxyl group and an amino group is preferable.

As the hydrophilic substance, a compound having at least one of a carboxy group and a sulfo group and at least one of a hydroxyl group and an amino group is preferable, and a compound represented by the following general formula (1) is more preferable.

(In the general formula (1), P represents at least one of a carboxy group and a sulfo group, A represents at least one of a hydroxyl group and an amino group, and R represents an m + n-valent linking group. Represents an integer of 3, n represents an integer of 1-3. The plurality of Ps and the plurality of As may be the same or different.)

In the general formula (1), R is preferably a linking group containing carbon, more preferably a linear or branched hydrocarbon group, and an alkylene group (for example, methylene) which is a divalent linking group. A group, an ethylene group, a propylene group, etc.), a phenylene group, etc. are more preferable, and an alkylene group is particularly preferable.
In the general formula (1), m is preferably 1 and n is preferably 1.

More specifically, the hydrophilizing substances include amino acids such as glycine and lysine, hydroxymethanesulfonic acid, hydroxyethanesulfonic acid, hydroxypropanesulfonic acid, hydroxybutanesulfonic acid, glycolic acid, lactic acid, tartronic acid, and glyceric acid. Examples thereof include hydroxybutyric acid, malic acid, tartaric acid, citramalic acid, citric acid, isocitrate, leucic acid, mevalonic acid, pantoic acid, aminomethanesulfonic acid, aminoethanesulfonic acid (taurine), aminopropanesulfonic acid and the like. The hydrophilic substance may be a salt, for example, an alkali metal salt (lithium salt, sodium salt).

<Joining process>
The method for producing a microchip of this embodiment includes a joining step of joining the pair of base materials after the immobilization step. The joining step provides a microchip in which both the hydrophilic and positively charged layers are imparted to the sampling section and the analysis section.

In the joining step, the first groove 16 in the chip component A and the region 17 in the chip component B face each other, and the region 19 in the chip component A and the second groove 18 in the chip component B are formed. The chip component A and the chip component B may be joined so as to face each other. As a result, the sampling unit 1 is formed at a position where the first groove 16 and the region 17 face each other, and the analysis unit 2 is formed at a position where the region 19 and the second groove 18 face each other.

The method of joining the pair of base materials (for example, the chip component A and the chip component B) is not particularly limited, and examples thereof include a method of heat-pressing the joint surfaces of the pair of base materials in contact with each other. ..

[Microchip]
The microchip according to one aspect of the present invention includes a sampling unit for collecting a sample and an analysis unit for analyzing the sample, and a cation in which a hydrophilic substance is bound to the sampling unit and the inner wall of the analysis unit. The sex polymer is immobilized. The microchip of this embodiment is a microchip in which both a hydrophilic and a positive charge layer are imparted to a sampling unit and an analysis unit, and can be produced, for example, by the method for producing a microchip of the present embodiment described above. Since the cationic polymer and the hydrophilizing substance used for producing the microchip of this embodiment are the same as those of the cationic polymer and the hydrophilizing substance described in the above-described method for producing the microchip of this embodiment, the description thereof will be described. Omit.

Further, in the microchip of this embodiment, from the viewpoint of suppressing the production cost, it is preferable that one unit constituting the microchip includes a sampling unit and an analysis unit. Such a microchip may be obtained by joining a pair of base materials as described in the method for producing a microchip described above, and the sampling unit and the analysis unit are a pair of bases. It may be a portion formed by joining the materials.

Further, it is preferable that the cationic polymer immobilized on the inner wall of the sampling section and the cationic polymer immobilized on the inner wall of the analysis section are the same, and the cationic polymer is immobilized on the inner wall of the sampling section. It is more preferable that the cationic polymer to which the hydrophilic substance is bound and the cationic polymer to which the hydrophilic substance is bound, which are immobilized on the inner wall of the analysis unit, are the same.

As an example, the microchip 100 shown in FIG. 3 will be described below. As shown in FIG. 3, the microchip 100 is formed by joining the chip component A and the chip component B, and has a sampling unit 1, an analysis unit 2, a sampling port 3, a sample reaching port 4, and a sample holding unit. A unit 5 and a running fluid holding unit 6 are provided. Hereinafter, each configuration will be described.

The sampling port 3 is a portion to which the sample to be analyzed is supplied (preferably spotted) from the opening, and is connected to the end portion of the sampling portion 1. The sample supplied to the sampling unit 1 is collected by the sampling unit 1.

The sampling unit 1 is a flow path for connecting the sample collection port 3 and the sample arrival port 4 and collecting the sample supplied to the sample collection port 3, preferably the sample collection port 3 using the capillary force. It is a microchannel for collecting the sample supplied to. The sample supplied to the sample collection port 3 is preferably collected by the sampling unit 1 by capillary force and sucked into the sample arrival port 4. At this time, since the sampling unit 1 is imparted with hydrophilicity, the sampling unit 1 is excellent in sample collection property, and the collected sample reaches the sample arrival port 4 in a short time through the sampling unit 1. do.

The size of the sampling unit 1 is not particularly limited, and for example, the depth is preferably 100 μm to 1000 μm, the width is 100 μm to 2000 μm, and the length is preferably 1 mm to 20 mm.

The sample arrival port 4 is connected to the end portion of the sampling unit 1 and is a portion for holding the sample reached through the sampling unit 1. Further, at the sample arrival port 4, for example, a diluted sample may be prepared by supplying a diluted solution through an opening and mixing the sample and the diluted solution. The main agent of the diluted solution is not particularly limited, and examples thereof include water and physiological saline, and preferred examples thereof include a liquid having a component similar to that of the running solution described later. Further, the diluent may be, for example, one in which a cathodic group-containing compound is added to the main agent. Examples of the cathode group-containing compound include cathode group-containing polysaccharides, and more specifically, sulfated polysaccharides, carboxylic oxide polysaccharides, sulfonated polysaccharides, phosphorylated polysaccharides and the like. As the carboxylic oxide polysaccharide, alginic acid and a salt thereof (for example, sodium alginate) are preferable. As the sulfated polysaccharide, for example, chondroitin sulfate is preferable. There are seven types of chondroitin sulfate, A, B, C, D, E, H, and K, and any of them may be used. The concentration of the cathodic group-containing compound (chondroitin sulfate) is preferably in the range of, for example, 0.01% by mass to 5% by mass.
The diluted sample obtained by mixing the sample with the diluent is supplied to the sample holding unit 5.

The sample holding unit 5 is a tank for holding the diluted sample supplied from the opening, and is connected to the end portion of the analysis unit 2.

The analysis unit 2 is a flow path for connecting the sample holding unit 5 and the running solution holding unit 6 and analyzing the sample (diluted sample) supplied to the sample holding unit 5, preferably the analysis of the sample by electrophoresis. It is a capillary tube that performs.

The size of the analysis unit 2 is not particularly limited, and for example, the depth is preferably 25 μm to 100 μm, the width is 25 μm to 100 μm, and the length is 5 mm to 150 mm.

The running solution holding unit 6 is connected to the end portion of the analysis unit 2 and is a tank for introducing the running solution supplied from the opening into the analysis unit 2. The running solution is a medium that is supplied from the opening of the running solution holding section 6 and introduced into the analysis section 2 to generate an electroosmotic flow in electrophoresis. The running solution is not particularly limited, but one using an acid is preferable. Examples of the acid include citric acid, maleic acid, tartaric acid, oxalic acid, fumaric acid, phthalic acid, malonic acid, and malic acid. In addition, the running solution preferably contains a weak base. Examples of the weak base include arginine, lysine, histidine, tris and the like. The pH of the running solution is preferably in the range of, for example, pH 4.5 to 6. Examples of the running solution buffer include MES, ADA, ACES, BES, MOPS, TES, HEPES and the like. Further, the cathodic group-containing compound described in the description of the diluent may be added to the running solution. The concentration of the cathodic group-containing compound (chondroitin sulfate or the like) is preferably in the range of, for example, 0.01% by mass to 5% by mass.

In the microchip 100, the diluted sample is supplied to the sample holding unit 5, the running solution is supplied to the running solution holding part 6, and the running solution is introduced into the analysis unit 2 by pressurization, and then the anode is attached to the sample holding part 5. The negative electrode is brought into contact with the running solution holding portion 6 to apply a voltage. As a result, an electric permeation flow is generated in the analysis unit 2, the diluted sample is introduced from the sample holding unit 5 to the analysis unit 2, and when the diluted sample moves from the sample holding unit 5 toward the running solution holding unit 6, the diluted sample is used. The components inside are separated. The voltage to be applied is, for example, preferably 0.5 kV to 10 kV, and more preferably 0.5 kV to 5 kV.

In the microchip 100, since the analysis unit 2 is provided with a positive charge layer, the analysis performance in the analysis unit 2 is excellent.

The component (substance to be analyzed) separated by the analysis unit 2 can be measured by an optical method such as absorbance measurement. For example, the absorbance may be measured by irradiating light from the detection window 15 on the chip component A side. When the substance to be analyzed contained in the diluted sample is hemoglobin, it is preferable to measure the absorbance at a wavelength of 415 nm, for example. Further, the component ratio in the diluted sample may be obtained by calculating the peak height, peak area, etc. of the electropherogram obtained by measuring the absorbance.

The position where the analysis unit 2 measures the separated components, that is, the position where the detection window 15 is arranged may be appropriately determined according to the length of the analysis unit 2 and the like.

〔kit〕
The microchip of this embodiment may be combined with a cartridge for storing a solution for analysis as a kit. The kit may include, for example, the microchip of the present embodiment and a cartridge for storing the diluent and the running solution, respectively.

[Analysis system]
Further, the microchip of this embodiment may be used as an analysis system in combination with an analyzer to which the microchip is attached. The analysis system includes, for example, the above-mentioned microchip 100 and an analyzer including an electrode, a light emitting unit, a measuring unit, a diluent tank, an electrophoresis liquid tank, a pump, a control unit, and the like.

Examples of the electrode include an anode inserted into the sample holding unit 5 and a negative electrode inserted into the running solution holding unit 6, which applies a predetermined voltage to the analysis unit 2.

The light emitting portion is, for example, a portion that emits light for measuring the absorbance and irradiates the detection window 15 with light. The light emitting unit includes, for example, an LED chip, an optical filter, a lens, or the like that emits light in a predetermined wavelength range. Further, the light emitting unit may be provided with a slit.

The measuring unit is, for example, a portion that receives the light emitted to the detection window 15 and measures the absorbance. The measuring unit includes, for example, a photodiode, a photo IC, and the like.

The diluent tank is, for example, a tank for storing the diluent supplied to the sample arrival port 4, and the running solution tank is, for example, a tank for storing the running solution supplied to the running solution holding unit 6.

The pump is, for example, a site for supplying the diluted solution to the sample arrival port 4 by applying pressure, supplying the running solution to the running solution holding unit 6 by applying pressure, and introducing the running solution into the analysis unit 2. Further, the diluted sample in the sample arrival port 4 may be supplied to the sample holding unit 5 by a pump.

The control unit controls each of the above-mentioned configurations in the analyzer, and includes, for example, a CPU, a memory, an interface, and the like.

Next, one aspect of the present invention will be described based on the following examples, but the present invention is not limited to the examples.

[Example 1]
<1: Preparation of chip parts>
PMMA (polymethyl methacrylate) was used as a material for producing the chip parts, and the chip parts A shown in FIG. 1 and the chip parts B shown in FIG. 2 were prepared by molding. The thickness of the chip parts A and B and the size of each configuration are as follows. By joining the chip component A and the chip component B in the subsequent process, as shown in FIG. 3, the sampling unit 1 formed from the first groove 16 and the region 17 on the surface of the chip component B, and the second A microchip 100 including a groove 18 and an analysis unit 2 formed from a region 19 on the back surface (joint surface) of the chip component A is formed.
(Chip part A)
Thickness: 1.5 mm
First groove 16 (sampling unit 1): depth 0.5 mm × width 1 mm × length 6 mm
Through hole 11 (sampling port 3): φ1.0 mm
Through hole 12 (sample arrival port 4): φ1.0 mm
Through hole 13 (sample holding portion 5) ... Capacity 10 μL
Through hole 14 (electrophoretic solution holding portion 6) ... Capacity 10 μL
(Chip part B)
Thickness: 1.5 mm
Second groove 18 ... Depth 0.04 mm x Width 0.04 mm x Length 30 mm

<2: Preparation of aqueous solution A>
Aqueous solution A was prepared by mixing the following components. In the aqueous solution A, the hydrophilic substance sodium hydroxymethanesulfonate is bound to the cationic polymer polydiallyldimethylammonium chloride. The hydrophilic substance (sodium hydroxymethanesulfonate) used in this example contains one sulfo group as an anionic functional group and one hydroxyl group as a hydrophilic functional group.
(Aqueous solution A)
-Polydialyldimethylammonium chloride (PDADMAC, polyquaternium-6, weight average molecular weight 100,000-500000, manufactured by Senka, trade name: FPA1000L) 1% w / v
・ Sodium hydroxide (NaOH, manufactured by Kishida Chemical Co., Ltd.) 10 mM
-Sodium hydroxymethanesulfonate 0.5 hydrate (HMS-Na, manufactured by Tokyo Chemical Industry Co., Ltd.) 100 mM

<3: Immobilization process>
Vacuum ultraviolet light (VUV) is applied to the joint surface of chip component A and chip component B using a VUV irradiation device (model number: MECL-3-500 manufactured by MD.com) to an integrated illuminance of 420 mJ / cm ² . And irradiated. Then, the chip component A and the chip component B were immersed in the above-mentioned aqueous solution A at room temperature (23 ± 2 ° C.) for 24 hours. After the immersion, the chip component A and the chip component B were thoroughly washed with water, and compressed air was injected to remove the water adhering to the chip component A and the chip component B. Through the series of steps up to this point, a cationic polymer to which hydroxymethanesulfonic acid is bound as a hydrophilic substance is immobilized on the bonding surface of the chip component A and the chip component B.

<4: Joining process>
With the joint surfaces of the chip component A and the chip component B in contact with each other, heat crimping (temperature 80 ° C., pressure 16 kgf / cm ² (1.57 MPa) for 1 minute) is performed to join the chip component A and the chip component B. bottom. As a result, as shown in FIG. 3, the sampling unit 1 and the analysis unit 2 on which the cationic polymer to which the hydrophilic substance was bound were immobilized produced the microchip 100 provided in the same unit.

<5: Hydrophilicity evaluation>
According to the following procedure, the hydrophilicity of the sampling part of the microchip was evaluated at two time points, immediately after production and after being sealed with a desiccant and stored at 50 ° C. for 1 week. Whole human blood was used as a sample.
1) A specified amount (3 μL) of sample was spotted on the sampling port of the sampling section. The spotted sample is sucked toward the sample arrival port by capillary force.
2) The time of instillation was set to 0 seconds, and the time required for the sample to reach the sample arrival port (time required for sample collection) was measured.
〔Evaluation criteria〕
Immediately after production and after storage at 50 ° C. for 1 week, the evaluation criteria for hydrophilicity are as follows. If it is evaluation A or evaluation B, the hydrophilicity of the sampling part is good.
A There was no difference in the time required for the sample to reach the sample arrival port immediately after production and after storage at 50 ° C. for 1 week.
B There was a difference in the time required for the sample to reach the sample arrival port immediately after production and after storage at 50 ° C for 1 week, but the sample was sample both immediately after production and after storage at 50 ° C for 1 week. You have reached the arrival port.
C The sample did not reach the sample arrival port immediately after production and at least one after storage at 50 ° C. for 1 week.
The results are shown in Tables 1 and 2 below.

<6: Positive charge layer function evaluation>
The positive charge layer function of the analysis unit in the microchip was evaluated by analyzing hemoglobin in whole human blood according to the following procedure. For the analysis, a special device made by ARKRAY for electrophoresis was used together with the microchip.
1) The microchip was set in a special device made by ARKRAY.
2) The following running solution (2) was added to the running solution holding tank (running solution holding part) of the microchip, and the running solution (2) was introduced into the analysis part by pressurization.
3) Human whole blood was diluted 41-fold with the following running solution (1), and the diluted sample was added to the sample holding tank (sample holding part).
4) The anode was brought into contact with the sample holding tank and the negative electrode was brought into contact with the running solution holding tank, and electrophoresis was started under 72 μA constant current control.
5) The absorbance at 415 nm was measured in the measuring section to obtain an electropherogram. Electrophoresis was performed for 60 seconds.
(Electrophoretic solution (1))
Citric acid 38 mM, chondroitin sulfate C sodium 0.95% w / v, 1- (3-sulfopropyl) pyridinium hydroxide intramolecular salt 475 mM, 2-morpholinoetan sulfonic acid 19 mM, Emargen LS-110 0.1% w / v, sodium azide 0.02% w / v, Proclin 300 0.025% w / v, dimethylaminoethanol (for pH adjustment), pH 6.0
[Electrophoretic solution (2)]
Citric acid 40 mM, chondroitin sulfate C sodium 1.25% w / v, piperazine 20 mM, emalgen LS-110 0.1% w / v, sodium azide 0.02% w / v, proclin 300 0.025% w / v, Dimethylaminoethanol (for pH adjustment), pH 5.0
〔Evaluation criteria〕
The evaluation criteria for the positive charge layer function are as follows.
A In the obtained electropherogram, it was confirmed that L-HbA1c (LA1C), S-HbA1c (s-A1C) and HbA0 were separated.
B In the obtained electropherogram, it could not be confirmed that L-HbA1c, S-HbA1c and HbA0 were separated.
The results are shown in FIG. 4 and Table 2 below.

[Example 2]
A microchip was produced in the same manner as in Example 1 except that the following aqueous solution B was prepared in place of the aqueous solution A in <2: Preparation of aqueous solution A> of Example 1. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 1. The hydrophilic substance (taurine) used in this example contains one sulfo group as an anionic functional group and one amino group as a hydrophilic functional group.
(Aqueous solution B)
-Polydialyldimethylammonium chloride (PDADMAC, polyquaternium-6, weight average molecular weight 100,000-500000, manufactured by Senka, trade name: FPA1000L) 1% w / v
・ Sodium hydroxide (NaOH, manufactured by Kishida Chemical Co., Ltd.) 10 mM
・ Taurine (manufactured by Nakarai) 100 mM

[Example 3]
A microchip was produced in the same manner as in Example 1 except that the following aqueous solution C was prepared in place of the aqueous solution A in <2: Preparation of aqueous solution A> of Example 1. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 1. The hydrophilic substance (lithium lactate) used in this example contains one carboxy group as an anionic functional group and one hydroxyl group as a hydrophilic functional group.
(Aqueous solution C)
-Polydialyldimethylammonium chloride (PDADMAC, polyquaternium-6, weight average molecular weight 100,000-500000, manufactured by Senka, trade name: FPA1000L) 1% w / v
・ Sodium hydroxide (NaOH, manufactured by Kishida Chemical Co., Ltd.) 10 mM
・ Lithium lactate (manufactured by Nakarai) 100 mM

[Example 4]
A microchip was produced in the same manner as in Example 1 except that the following aqueous solution D was prepared in place of the aqueous solution A in <2: Preparation of aqueous solution A> of Example 1. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 1. The hydrophilic substance (glycine) used in this example contains one carboxy group as an anionic functional group and one amino group as a hydrophilic functional group.
(Aqueous solution D)
-Polydialyldimethylammonium chloride (PDADMAC, polyquaternium-6, weight average molecular weight 100,000-500000, manufactured by Senka, trade name: FPA1000L) 1% w / v
・ Sodium hydroxide (NaOH, manufactured by Kishida Chemical Co., Ltd.) 10 mM
・ Glycine (manufactured by Nakarai) 100 mM

[Example 5]
A microchip was produced in the same manner as in Example 1 except that the following aqueous solution E was prepared in place of the aqueous solution A in <2: Preparation of aqueous solution A> of Example 1. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 1. The hydrophilic substance (lysine) used in this example contains one carboxy group as an anionic functional group and two amino groups as a hydrophilic functional group.
(Aqueous solution E)
-Polydialyldimethylammonium chloride (PDADMAC, polyquaternium-6, weight average molecular weight 100,000-500000, manufactured by Senka, trade name: FPA1000L) 1% w / v
・ Sodium hydroxide (NaOH, manufactured by Kishida Chemical Co., Ltd.) 10 mM
・ Lysine (manufactured by Nakarai) 100 mM

[Example 6]
Except that the following aqueous solutions F and G were prepared in place of the aqueous solution A in <2: Preparation of aqueous solution A> of Example 1 and the following operations were performed in <3: Immobilization step>. A microchip was produced in the same manner as in Example 1. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 1.
(Aqueous solution F)
-Polydialyldimethylammonium chloride (PDADMAC, polyquaternium-6, weight average molecular weight 100,000 to 500,000, manufactured by Senka, trade name: FPA1000L) 1% w / v
・ Sodium hydroxide (NaOH, manufactured by Kishida Chemical Co., Ltd.) 10 mM
(Aqueous solution G)
-Sodium hydroxymethanesulfonate 0.5 hydrate (HMS-Na, manufactured by Tokyo Chemical Industry Co., Ltd.) 100 mM

<3: Immobilization process (modification process)>
Vacuum ultraviolet light (VUV) is applied to the joint surface of chip component A and chip component B using a VUV irradiation device (model number: MECL-3-500 manufactured by MD.com) to an integrated illuminance of 420 mJ / cm ² . And irradiated. Then, the chip component A and the chip component B were immersed in the above-mentioned aqueous solution F at room temperature (23 ± 2 ° C.) for 24 hours. After the immersion, the chip component A and the chip component B were thoroughly washed with water, and compressed air was injected to remove the water adhering to the chip component A and the chip component B. By this step, the cationic polymer is immobilized on the joint surface of the chip component A and the chip component B.

<3: Immobilization step (hydrophilization step)>
The chip component A and the chip component B were immersed in the above-mentioned aqueous solution G at room temperature (23 ± 2 ° C.) for 10 minutes. After the immersion, the chip component A and the chip component B were thoroughly washed with water, and compressed air was injected to remove the water adhering to the chip component A and the chip component B. By this step, the hydrophilic substance is bonded to the cationic polymer immobilized on the joint surface of the chip component A and the chip component B, and the hydrophilic step is completed. In this example, sodium hydroxymethanesulfonate was used as the hydrophilizing substance as in Example 1.

[Example 7]
Example 6 except that the following aqueous solution J was prepared in place of the aqueous solution G in Example 6 and the aqueous solution J was used in place of the aqueous solution G in <3: Immobilization step (hydrophilization step)>. The microchip was manufactured in the same manner as in the above. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 6. In this example, trisodium citrate containing three carboxy groups as an anionic functional group and one hydroxyl group as a hydrophilic functional group was used as the hydrophilic substance.
(Aqueous solution J)
・ Trisodium citrate (manufactured by Wako Pure Chemical Industries, Ltd.) 100 mM

[Comparative Example 1]
A microchip was produced in the same manner as in Example 6 except that <3: Immobilization step (hydrophilization step)> was not carried out in Example 6. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 6.

[Comparative Example 2]
In Example 6, the following aqueous solution H was prepared in place of the aqueous solution G, <3: immobilization step (modification step)> was carried out, and <3: immobilization step (hydrophilization step)> was performed on the aqueous solution G. A microchip was produced in the same manner as in Example 6 except that the aqueous solution H was used instead of the above. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 6. In this comparative example, silica (a hydrophilic substance that cannot be bonded to a cationic polymer), which is fine particles of an inorganic compound, was used as the hydrophilic substance.
(Aqueous solution H)
・ Silica-containing aqueous solution (main component: silicon dioxide, trade name: cell face coat, manufactured by Marusho Sangyo Co., Ltd.)

[Comparative Example 3]
In Example 6, the following aqueous solution I was prepared in place of the aqueous solution G, <3: immobilization step (modification step)> was carried out, and <3: immobilization step (hydrophilization step)> of the aqueous solution G. A microchip was produced in the same manner as in Example 6 except that the aqueous solution I was used instead. With respect to the manufactured microchip, <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> were carried out in the same manner as in Example 6. In this comparative example, a polyoxyalkylene alkyl ether (a hydrophilic substance that cannot be bonded to a cationic polymer) that does not contain an anionic functional group and contains a polyoxyalkylene chain as a hydrophilic functional group is used as the hydrophilic substance. used.
(Aqueous solution I)
-Polyoxyalkylene alkyl ether (trade name: Emargen LS-110, manufactured by Kao Corporation) 0.1% w / v

The results of <5: hydrophilicity evaluation> and <6: positive charge layer function evaluation> for Examples 2 to 7 and Comparative Examples 1 to 3 are shown in Tables 1 and 2 below.

As shown in Tables 1 and 2, in Examples 1 to 7 (particularly Examples 1 to 6), the hydrophilicity was good both immediately after production and after storage at 50 ° C. for 1 week. On the other hand, in Comparative Examples 1 and 3, the hydrophilicity was poor immediately after production and at least one after storage at 50 ° C. for 1 week.
Further, as shown in FIG. 4, it was confirmed that L-HbA1c, S-HbA1c and HbA0 were separated in Examples 1 to 7, and the positive charge layer function evaluation was good. On the other hand, in Comparative Examples 2 and 3, it could not be confirmed that L-HbA1c, S-HbA1c and HbA0 were separated, and the positive charge layer function evaluation was poor.
As described above, in Examples 1 to 7, the two functions of the hydrophilic and positive charge layers were compatible with each other, whereas in Comparative Examples 1 to 3, at least one of the functions of the hydrophilic and positive charge layers was poor. Yes, the two functions of hydrophilicity and positive charge layer were not compatible.

1 Sample collection unit, 2 Analysis unit, 3 Sample collection port, 4 Sample arrival port, 5 Sample holding part, 6 Running fluid holding part, 11, 12, 13, 14 Through holes, 15 Detection window, 16 First groove (Region where the sampling part is formed), 17 area (Region where the sampling part is formed), 18 Second groove (Region where the analysis part is formed), 19 area (Region where the analysis part is formed) , A, B chip parts, 100 microchips

Claims (17)

A sampling port where a sample containing an analytical component is spotted, and
A first flow path into which the sample spotted on the sampling port is introduced, and
A second flow path that is provided apart from the first flow path and that analyzes the sample introduced into the first flow path, and a second flow path that analyzes the sample.
A microchip in which a cationic polymer in which a hydrophilic substance is bound is immobilized on the inner walls of the first flow path and the second flow path. The microchip according to claim 1, wherein one unit includes the first flow path and the second flow path. The microchip according to claim 1 or 2, wherein the first flow path and the second flow path are sites formed by joining a pair of base materials. Claims 1 to claim that the cationic polymer immobilized on the inner wall of the first flow path and the cationic polymer immobilized on the inner wall of the second flow path are the same. The microchip according to any one of 3. The microchip according to any one of claims 1 to 4, wherein the hydrophilic substance has one or more anionic functional groups. The microchip according to claim 5, wherein the anionic functional group contains at least one of a carboxy group and a sulfo group. The microchip according to any one of claims 1 to 6, wherein the hydrophilic substance has one or more hydrophilic functional groups. The microchip according to claim 7, wherein the hydrophilic functional group contains at least one of a hydroxyl group and an amino group. The microchip according to any one of claims 1 to 8, wherein the cationic polymer contains at least one of polyquaternium and a dimethylamine-epichlorohydrin copolymer. The microchip according to any one of claims 1 to 9, wherein the first flow path uses a capillary force to collect the sample. The microchip according to any one of claims 1 to 10, wherein the second flow path is a capillary tube. The microchip according to any one of claims 1 to 11.
An analyzer to which the microchip is attached and
Analytical system with. Sample containing analyte is have you a pair of substrates provided with a sampling port to be spotted, region and the first first flow path spotted by the sample in the sample collection port is introduced is formed Cationicity in which a hydrophilic substance is bound to a region where a second flow path is formed to analyze the sample introduced into the first flow path, which is provided apart from the region where the first flow path is formed. The immobilization process for immobilizing the polymer and
The first flow path and the first flow path, which includes a joining step of joining the pair of substrates after the immobilization step, and the cationic polymer to which the hydrophilized substance is bound are immobilized by the joining step. A method for manufacturing a microchip in which the second flow path is formed. The fixing step is a modified step of fixing the said cationic polymer in a region where the first region and the second flow path is formed in the flow path is formed, immobilized the cationic polymer The hydrophilization step of binding the hydrophilization substance to the
13. The method for manufacturing a microchip according to claim 13. In the immobilization step, after the hydrophilic substance is bound to the cationic polymer, the hydrophilic substance is formed in the region where the first flow path is formed and the region where the second flow path is formed. The method for producing a microchip according to claim 13, wherein the bonded cationic polymer is immobilized. A sampling unit that sucks and collects samples by capillary force,
A capillary tube for analyzing the sample is provided.
A microchip in which a cationic polymer in which a hydrophilic substance is bound is immobilized on the sampling section and the inner wall of the capillary tube. An immobilization step of immobilizing a cationic polymer in which a hydrophilic substance is bound to a region where a sampling part for collecting a sample is formed and a region where an analysis part for analyzing the sample is formed on a pair of base materials.
The sampling section and the analysis, which include a joining step of joining the pair of substrates after the immobilization step, in which the cationic polymer to which the hydrophilic substance is bound is immobilized by the joining step. Part is formed,
The sampling unit sucks and collects a sample by capillary force, and collects the sample.
The analysis unit is a method for manufacturing a microchip which is a capillary tube .

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