JP5237972B2 - Body fluid component analyzer - Google Patents

Description

  The present invention relates to an analytical instrument for measuring a body fluid component of a living body.

  As a body fluid component analysis device, a technique for analyzing a body fluid component by dropping a small amount of a body fluid sample onto a test piece holding a reagent and measuring optical characteristics after the sample reacts with the reagent is known. (See Patent Documents 1 and 2). This analytical instrument supplies a sample of body fluid to a sample supply port provided in a test piece, and then transfers the sample to a measurement chamber equipped with a reagent through a flow path to generate a color reaction in the measurement chamber. Thus, a desired body fluid component can be measured by measuring the absorbance in the measurement chamber. In addition, when the sample to be applied is whole blood but the measurement target is a component in plasma, etc., a filtering means is provided as appropriate, and blood cell components in the whole blood sample are separated and filtered, and then placed in the measurement chamber. It can also be configured to be transported.

  The analytical instrument is configured by stacking a plurality of plate materials, and a flow path from the sample supply port to the measurement chamber is formed by a groove or the like provided in a part of the layers. The sample is transferred from the sample supply port to the measurement chamber by suction using a pump or the like or pressurization from the sample supply port.

  At this time, if the layer constituting the flow path and the measurement chamber is an impermeable and breathable plate, the sample can be transferred to the measurement chamber without providing an air vent or the like at the tip of the measurement chamber. Further, since there is no air vent hole at the end of the measurement chamber, there is no possibility that the sample passes through the measurement chamber and leaks to the outside.

  A method of processing and using a polytetrafluoroethylene (hereinafter referred to as PTFE) membrane as a plate member having impermeability and air permeability is known. Here, the PTFE film has many variations in film thickness and has the property that deformation caused by processing such as lamination is difficult to restore.

  As described above, the analytical instrument has a laminated structure, and the PTFE membrane is sandwiched between plastic plates to form a flow path. In the measurement chamber, the plastic plate is formed to be transparent, and the intensity of incident light and transmitted light of the part can be measured, and the absorbance can be measured therefrom.

  At this time, when measuring the absorbance, the light incident on the measurement unit is measured as transmitted light after passing through the plurality of stacked plate members one after another. That is, the intensity of transmitted light varies depending on the plate thickness, and the measured value of absorbance also varies. Here, if the film thickness of the PTFE film is different for each individual analytical instrument, the measurement results also vary, leading to a decrease in accuracy.

  On the other hand, when cellulose acetate or cellulose mixed ester is used as the impervious and breathable plate member, these materials have good recovery from deformation caused by processing. The variation in thickness is small, and therefore the variation in measurement results is slight. However, unlike PTFE, cellulose acetate and cellulose mixed ester do not inherently have water impermeability, and therefore need to be separately subjected to water / oil repellent treatment. As the water / oil repellent treatment, a fluorine-based surface treatment is generally used.

However, a conventional water repellent treatment agent usually contains a surfactant in order to improve the dispersibility of the fluorine compound. Therefore, when a porous plate member such as cellulose acetate or cellulose mixed ester is treated with such a treatment agent, the sample in which the reagent is dissolved in the porous plate member soaks due to such a surfactant. There was a fault that would Further, there are cases where a surfactant is contained in the reagent, and in this case, it has been confirmed that the same phenomenon occurs. When this phenomenon occurs, the apparent absorbance is lowered, so that the measured value is not stable and the measurement accuracy is lowered.
JP-A-8-114539 JP 2002-2224090 A

  Therefore, it is a case where a specific component such as total cholesterol in body fluid is analyzed, and even when the reagent contains a surfactant, there is no penetration of the sample in which the reagent is dissolved in the porous plate member. There has been a demand for a highly accurate analytical instrument that can measure absorbance and has little variation among individuals.

The present invention has been made to solve the above problems, and the invention according to claim 1 is directed to a sample supply port through which a sample is supplied, a measurement chamber in which the sample is measured, and the sample A flow path communicating with the supply port and the measurement chamber, the flow path and the measurement chamber are formed by stacking a plurality of plate members, and a plate forming at least the flow path among the plurality of plate members is A porous plate member having air permeability, wherein the surface of the porous plate member having air permeability has at least a substance 1 and a siloxane group mainly composed of a fluorocarbon group, a hydrocarbon group, and a silyl group. A body fluid component analyzing instrument characterized in that it is covered with a water- and oil-repellent composite film containing a substance 2 as a main component and chemically bonded to the surface. Here, the mixing composition ratio of the substance 1 mainly composed of a fluorocarbon group, a hydrocarbon group and a silyl group and the substance 2 mainly composed of a siloxane group is adjusted according to the required water and oil repellency. While possible, a range of 1:10 to 1: 0 is preferred, but a range of 1: 3 to 1: 0 is more preferred.

  According to the first aspect of the present invention, since the porous plate member forming the flow path has water permeability and oil repellency while having air permeability, the sample in which the reagent is dissolved in the porous plate member is not soaked. It is possible to provide a body fluid component analyzer that can transfer a sample applied to a sample supply port to a measurement chamber by an operation such as pressurization or suction without providing an air vent hole. Further, since there is no air vent hole, the sample does not leak outside through the measurement chamber. Furthermore, since the water and oil repellency is high and reliable, even a sample containing a surfactant is stable without being soaked into the porous plate member having air permeability over time. Measurement is possible.

  According to a second aspect of the present invention, there is provided the body fluid component analyzing instrument according to the first aspect, wherein the material of the porous plate member having air permeability is cellulose acetate or cellulose mixed ester.

  According to invention of Claim 2, since the porous plate member which has air permeability which is a component of a laminated structure consists of cellulose acetate or a cellulose mixed ester, its reactivity with a composite film formation solution is high, A water- and oil-repellent composite film that is an ultra-thin film of nanometer level and has high peel resistance can be formed on the surface. Therefore, even if the porous plate member having air permeability forms a water- and oil-repellent film, the vent hole is hardly blocked. Therefore, compared with the conventional fluorine-based water / oil repellent treatment, the air permeability is high and the water / oil repellency can be kept high. Moreover, since the variation in the plate thickness for each analytical instrument is small, it is possible to provide an analytical instrument for a body fluid component with small variations in measurement results.

  According to a third aspect of the present invention, the measurement chamber is provided with a reagent that reacts with a specific component in a body fluid and colors, and the reagent includes a surfactant. It is a device for analyzing body fluid components.

  According to the invention of claim 3, while the effect of the invention of claim 2 is obtained, the reagent provided in the measurement chamber reacts with a specific component in the body fluid to cause a color reaction. Due to the surfactant contained in the reagent, the reagent dissolves in the sample and the surface tension decreases. Here, if the sample in which the reagent is dissolved soaks into the peripheral member, a coloration different from the color that should be originally observed is measured. However, since the porous plate member having air permeability has high water and oil repellency, it does not penetrate even in a sample containing a surfactant, and it is an instrument for analyzing a body fluid component that has a stable color reaction. Can provide.

The invention described in claim 4 includes a sample supply port through which a sample is supplied, a measurement chamber in which measurement of the sample is performed, and a flow path that connects the sample supply port and the measurement chamber. The flow channel and the measurement chamber are formed by laminating a plurality of plate members, and at least a plate forming the flow channel among the plurality of plate members is a porous plate member having air permeability, and is a body fluid component analysis instrument In the production method of the above, the porous plate member having air permeability is made of at least a substance mainly composed of a fluorocarbon group, a hydrocarbon group and a chlorosilyl group, and a substance or an alkoxysilyl group mainly composed of a chlorosilyl group. A method for producing a body fluid component analysis instrument, comprising a step of contacting a composite film forming solution containing a substance containing as a main component and a non-aqueous organic solvent. Here, the mixing composition ratio of the substance mainly composed of fluorocarbon group, hydrocarbon group and chlorosilyl group and the substance mainly composed of chlorosilyl group or the substance mainly composed of alkoxysilyl group is required. Although it can be adjusted according to the water / oil repellency, the range of 1:10 to 1: 0 is preferable, but the range of 1: 3 to 1: 0 is more preferable.

  According to the invention described in claim 4, the body fluid component analysis instrument according to claim 1 can be manufactured.

The invention according to claim 5 includes a sample supply port through which a sample is supplied, a measurement chamber in which measurement of the sample is performed, and a flow path that connects the sample supply port and the measurement chamber. The flow channel and the measurement chamber are formed by laminating a plurality of plate members, and at least a plate forming the flow channel among the plurality of plate members is a porous plate member having air permeability. In the production method, the porous plate member having air permeability, a substance mainly containing at least a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group, a substance containing a silanol condensation catalyst or a chlorosilyl group, A method for producing a body fluid component analytical instrument comprising a step of contacting a composite film forming solution containing a non-aqueous organic solvent. Here, the mixing composition ratio of the substance mainly composed of fluorocarbon group, hydrocarbon group and alkoxysilyl group and the substance containing chlorosilyl group can be adjusted according to the required water and oil repellency. However, the range of 1:10 to 1: 0 is preferable, but the range of 1: 3 to 1: 0 is more preferable.

  According to the invention described in claim 5, since the porous plate member forming the flow path has water permeability and oil repellency while having air permeability, the sample in which the reagent is dissolved in the porous plate member is not soaked. It is possible to manufacture a body fluid component analyzer that can transfer a sample applied to a sample supply port to a measurement chamber by an operation such as pressurization or suction without providing an air vent hole. . Further, since there is no air vent hole, the sample does not leak outside through the measurement chamber. Furthermore, since the water and oil repellency is high and reliable, even a sample containing a surfactant is stable without being soaked into the porous plate member having air permeability over time. Measurement is possible.

The invention described in claim 6 includes a sample supply port through which a sample is supplied, a measurement chamber in which measurement of the sample is performed, and a flow path that connects the sample supply port and the measurement chamber. The flow channel and the measurement chamber are formed by laminating a plurality of plate members, and at least a plate forming the flow channel among the plurality of plate members is a porous plate member having air permeability. In the production method of the porous plate member having air permeability, a substance having at least a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group as main components, a substance having an alkoxysilyl group as main components, An apparatus for analyzing a body fluid component, comprising a step of contacting a silanol condensation catalyst or a substance containing a chlorosilyl group and a composite film forming solution containing a non-aqueous organic solvent. It is a production method. Here, the mixing composition ratio of the substance mainly composed of a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group and the substance mainly composed of the alkoxysilyl group or the substance containing the chlorosilyl group is required. Although it can be adjusted according to the water and oil repellency, the range of 1:10 to 1: 0 is preferable, but the range of 1: 3 to 1: 0 is more preferable.

  According to the sixth aspect of the present invention, the body fluid component analysis instrument according to the first aspect can be manufactured.

  Invention of Claim 7 is obtained by the manufacturing method in any one of Claims 4-6, The material of the said porous plate member which has air permeability is a cellulose acetate or a cellulose mixed ester, The said measurement An analysis device for a body fluid component, wherein the chamber is provided with a reagent that reacts with a specific component in the body fluid to develop a color, and the reagent contains a surfactant.

  According to the seventh aspect of the present invention, a water- and oil-repellent coating that is a nanometer level ultra-thin film and has high peel resistance is formed on the surface of the porous plate member. The quality plate member can maintain high water and oil repellency while maintaining air permeability. Moreover, since the variation in the plate thickness for each analytical instrument is small, it is possible to provide an analytical instrument for a body fluid component with small variations in measurement results. In addition, the porous plate member with air permeability has high water and oil repellency, so even if the sample contains a surfactant, it does not penetrate and provides an analytical instrument for body fluid components with a stable color reaction. can do.

  According to an eighth aspect of the present invention, there is provided a step of supplying a sample to the sample supply port using the body fluid component analyzing instrument according to the third or seventh aspect, and the sample is introduced into the measurement chamber through the flow path. And a step of measuring absorbance in the measurement chamber after the sample and the color sample have undergone a color reaction, and a method for analyzing a body fluid component.

  According to the invention described in claim 8, the variation of each analytical instrument can be ignored, and a highly efficient and highly reliable analysis result can be obtained.

  According to the present invention, since the plate member forming the flow channel has air permeability and water and oil repellency, it is applied to the sample supply port by an operation such as pressurization or suction without providing an air vent. It is possible to provide a body fluid component analyzing instrument capable of transferring a sample to a measurement chamber. Further, since there is no air vent hole, the sample does not leak outside through the measurement chamber. Furthermore, since the water and oil repellency is high and reliable, the sample does not penetrate into the porous plate member having air permeability even after a lapse of time, and stable measurement is possible.

It is sectional drawing which shows the analytical instrument which concerns on embodiment of this invention. It is a disassembled perspective view which shows the analytical instrument which concerns on embodiment of this invention. It is a graph which shows the time change of the light absorbency measured value in the Example and comparative example of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 1st plate 2 2nd plate 3 3rd plate 4 Sample supply port 5 Flow path 51 Through-hole 52 Groove 6 Measurement chamber 61 Through-hole 62 Reagent holding part

  Next, embodiments of the present invention will be described with reference to the drawings.

  First, FIG. 1 and FIG. 2 show an embodiment of an analytical instrument according to the present invention. 1 is a cross-sectional view, and FIG. 2 is an exploded perspective view. In this embodiment, a plasma sample is applied to measure total cholesterol. Note that whole blood can be applied as a sample if a separate filtration means is provided on the sample supply port so that blood cell components can be supplied after being separated and filtered. Furthermore, if the filtering means contains an agglutinating reagent that reacts and aggregates with lipoproteins other than high-density lipoprotein (HDL), and the aggregate can be separated and filtered by the filtering means, whole blood By applying a sample, an analytical instrument capable of selectively quantifying cholesterol in high density lipoprotein (HDL-C) can be realized.

  As shown in FIG. 1 and FIG. 2, the analytical instrument according to the present embodiment includes a first plate 1, a second plate 2, and a third plate 3 via an adhesive layer (not shown) such as a double-sided tape. It is composed by bonding.

  The first plate 1 is provided with a sample supply port 4 composed of a through hole. The second plate 2 is provided with a through hole 51 at a position substantially concentric with the sample supply port 4, and a through hole 61 is provided at a distance from the second plate 2, and the groove 52 communicates with the two through holes 51, 61. . The third plate 3 is provided with a reagent holding part 62 substantially concentrically with the through hole 61 to hold a reagent (not shown). The reagent holding part 62 can be formed by forming a recess at the position of the third plate 3 and filling the reagent therein, but it can also be formed by attaching a reagent to a flat part. The through hole 51 and the groove 52 constitute the flow path 5, and the through hole 61 and the reagent holding part 62 constitute the measurement chamber 6.

  For the first plate 1 and the third plate 3, a plastic material such as polyethylene terephthalate (PET) or AS resin is preferable because it can be easily processed. However, the present invention is not limited to this as long as a flow path in which the sample does not leak can be formed.

  If a porous material that is impermeable and breathable is used for the second plate 2, the sample can be transferred to the measurement chamber 3 without providing an air vent or the like. Here, when a material that does not inherently impervious water is applied to the second plate 2, it is necessary to separately perform water and oil repellent treatment in order to impart impermeability while maintaining air permeability.

  In addition, it is easy to process the second plate 2 by processing filter paper or the like. It can be produced by processing a commercially available PTFE membrane or a filter paper made of a commercially available cellulose acetate or cellulose mixed ester.

  Of these, PTFE is inherently water-impermeable and is advantageous in that it does not require water / oil repellent treatment. However, there is a large variation in film thickness, and the property that the film thickness is likely to change due to processing such as stacking and is difficult to restore, which causes variations in absorbance measurement results.

  On the other hand, cellulose acetate or cellulose mixed ester is advantageous in ensuring the reproducibility of the absorbance measurement results because the variation in film thickness is small and the change in film thickness caused by processing is small. It is not water-based. Therefore, when using as the 2nd plate 2, it is necessary to perform a water- and oil-repellent treatment separately.

  The surface treatment using a fluorine-based oligomer is generally used for the water / oil repellent treatment, but the surface treatment using a fluorine-based oligomer impairs air permeability because the film thickness is thick. On the other hand, in the embodiment of the present invention, a repellent film composed of a composite film containing at least a substance 1 mainly composed of a fluorocarbon group, a hydrocarbon group and a silyl group and a substance 2 mainly composed of a siloxane group. By forming a film with a nano-level film thickness having water / oil repellency, water / oil repellency is imparted without impairing air permeability. The specific procedure of the water / oil repellent treatment of the present invention will be described below.

(Embodiment 1)
A composite film comprising at least a substance mainly composed of a fluorocarbon group, a hydrocarbon group and a chlorosilyl group, a substance mainly composed of a chlorosilyl group or a substance mainly composed of an alkoxysilyl group, and a non-aqueous organic solvent. The forming solution is adjusted in an airtight container and in an atmosphere with a humidity of 30% or less (in N ₂ gas or dry air, but a humidity of 5% or less is more preferable). Next, in the same atmosphere, an appropriate amount of the treatment solution is placed in a stainless steel vat and a porous membrane (either cellulose acetate or cellulose mixed ester is acceptable) is immersed in the treatment solution (because it is a solid-liquid interface reaction, it is porous. The membrane may be brought into contact with the composite film forming solution). At this time, care should be taken to ensure that the solution is immersed in the solution without air bubbles being mixed in or one side contacting the bat. Although there is no strict regulation on the immersion time, the current time is about 30 minutes.

  Next, after removing the porous membrane from the container, the porous membrane is left to dry in the same atmosphere. Although it depends on the type of solvent, in the case of a fluorocarbon organic solvent, the solvent volatilizes within a few minutes. At present, two of the four corners of the porous membrane (sheets) are “suspended” and dried. Here, the reason why the treatment is performed in an airtight container and in an atmosphere having a humidity of 30% or less is that when a film is formed, a hydrochloric acid gas harmful to the human body is generated due to a dehydrochlorination reaction.

  Finally, the dried porous membrane is taken out in ordinary air and left in the room overnight. By leaving it to stand overnight, the unreacted chlorosilyl group remaining on the surface of the porous membrane completely reacts with moisture in the air, and a predetermined water / oil repellency is obtained.

  In this method, the porous film made of at least cellulose acetate and cellulose mixed ester contains active hydrogen on the surface, and therefore the main component is a substance mainly composed of a fluorocarbon group, a hydrocarbon group, and a chlorosilyl group, and a chlorosilyl group. Dehydrochlorination reaction with a chlorosilyl group of a substance having an alkoxysilyl group as a main component and a substance 1 having a fluorocarbon group, a hydrocarbon group and a silyl group as main components and a siloxane group as a main component A film having a water and oil repellency formed of a composite film containing the substance 2 to be formed is chemically bonded to the surface of the porous film with a film thickness of nanometer level. Since the film thickness formed here is on the nanometer level, the air permeability of the porous film is hardly impaired. Even when a substance mainly composed of an alkoxysilyl group is used instead of a substance mainly composed of a chlorosilyl group, the generated hydrochloric acid has a catalytic action, and thus chemically bonds in the same manner.

  At this time, the mixing composition ratio of the substance 1 mainly composed of a fluorocarbon group, a hydrocarbon group and a silyl group and the substance 2 mainly composed of a siloxane group is adjusted according to the required water and oil repellency. While possible, a range of 1:10 to 1: 0 is preferred, but a range of 1: 3 to 1: 0 is more preferred.

(Embodiment 2)
A composite film forming solution containing at least a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group as a main component, a silanol condensation catalyst or a substance containing a chlorosilyl group, and a non-aqueous organic solvent has a humidity of 30% or less. atmosphere ₍₂ gas N, or may be dry air, but more preferred is that if the humidity 5%) to adjust. Next, in normal air, an appropriate amount of the treatment solution is placed in a stainless steel vat, and a porous membrane (either cellulose acetate or cellulose mixed ester is acceptable) is immersed in the treatment solution (since it is a solid-liquid interface reaction, The porous film may be brought into contact with the composite film forming solution). At this time, care should be taken to ensure that the solution is immersed in the solution without air bubbles being mixed in or one side contacting the bat. Although there is no strict regulation on the immersion time, the current time is about 1 hour.

  Next, after removing the porous membrane from the container, the porous membrane is left to dry in the same atmosphere. Although depending on the type of solvent, in the case of a fluorocarbon organic solvent, the solvent volatilizes within a few minutes. At present, two of the four corners of the porous membrane (sheets) are “suspended” and dried. In addition, since the film formation in this case is a dealcoholization reaction, there is no problem even if the treatment is performed in normal air.

  Finally, the dried porous membrane is left in the room overnight. By leaving it overnight, the hydrolysis reaction between the unreacted drug remaining on the surface of the porous membrane and the moisture in the air proceeds, and a predetermined water and oil repellency is obtained.

  In this method, since the porous film made of at least cellulose acetate and cellulose mixed ester contains active hydrogen on the surface, the alkoxysilyl group and silanol of a substance mainly composed of a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group. A film having a water- and oil-repellent property containing a substance 1 mainly composed of at least a fluorocarbon group, a hydrocarbon group, and a silyl group and having a water- and oil-repellent property is porous with a film thickness of nanometer level. It is formed by chemically bonding to the surface of the membrane. Since the film thickness formed here is on the nanometer level, the air permeability of the porous film is hardly impaired.

(Embodiment 3)
A substance mainly comprising at least a fluorocarbon group, a hydrocarbon group and an alkoxysilyl group; a substance mainly comprising an alkoxysilyl group; a substance containing a silanol condensation catalyst or a chlorosilyl group; and a non-aqueous organic solvent. composite film-forming solution 30% in less ambient humidity comprising (N ₂ gas, or may be dry air, but more preferred is that if the humidity 5%) to adjust. Next, in normal air, an appropriate amount of the treatment solution is placed in a stainless steel vat, and a porous membrane (either cellulose acetate or cellulose mixed ester is acceptable) is immersed in the treatment solution (since it is a solid-liquid interface reaction, The porous film may be brought into contact with the composite film forming solution). At this time, care should be taken to ensure that the solution is immersed in the solution without air bubbles being mixed in or one side contacting the bat. Although there is no strict regulation on the immersion time, the current time is about 1 hour.

  Next, after removing the porous membrane from the container, the porous membrane is left to dry in the same atmosphere. Although depending on the type of solvent, in the case of a fluorocarbon organic solvent, the solvent volatilizes within a few minutes. At present, two of the four corners of the porous membrane (sheets) are “suspended” and dried. In addition, since the film formation in this case is a dealcoholization reaction, there is no problem even if the treatment is performed in normal air.

  Finally, the dried porous membrane is left in the room overnight. By leaving it overnight, the hydrolysis reaction between the unreacted drug remaining on the surface of the porous membrane and the moisture in the air proceeds, and a predetermined water and oil repellency is obtained.

  In this method, since the porous membrane comprising at least cellulose acetate and cellulose mixed ester contains active hydrogen on the surface, a substance mainly composed of a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group and an alkoxysilyl group are contained. Substance 1 mainly composed of fluorocarbon group, hydrocarbon group and silyl group and substance mainly composed of siloxane group by dealcoholization reaction in the presence of silanol catalyst and alkoxysilyl group of substance mainly composed of substance 2 is formed by chemically bonding to the surface of the porous film with a thickness of nanometer level. Since the film thickness formed here is on the nanometer level, the air permeability of the porous film is hardly impaired.

  At this time, the mixing composition ratio of the substance 1 mainly composed of a fluorocarbon group, a hydrocarbon group and a silyl group and the substance 2 mainly composed of a siloxane group is adjusted according to the required water and oil repellency. While possible, a range of 1:10 to 1: 0 is preferred, but a range of 1: 3 to 1: 0 is more preferred.

  In addition, it is described in Unexamined-Japanese-Patent No. 2005-206790 that according to this process, compared with Embodiment 2, the coating film which has higher density and durability and water / oil repellency is formed.

  Next, the third plate 3 will be described. On the third plate 3, a reagent (not shown) is applied and dried in the reagent holding unit 62 and held. In the present embodiment, the measurement target is total cholesterol, and therefore, the reagent shown below is used as the reagent. However, the reagent is not limited to this adjustment reagent. In addition, the following density | concentration is a density | concentration at the time of application | coating.

Reagent preparation
Cholesterol esterase (CE) 200 U / mL
Cholesterol dehydrogenase (CHDH) 150 U / mL
Diaphorase (DI) 200 U / mL
Nicotinamide adenine dinucleotide (NAD) 20 mM
Water-soluble tetrazolium salt (WST-4) 60 mM
Surfactant (Triton (registered trademark) X-100) 1.0% w / v
Good buffer (TAPS) pH 8.5 200 mM

  Next, the analysis procedure using the instrument according to this embodiment and the function of the instrument will be described.

  A sample is dropped into the sample supply port 4. Since the flow channel has water and oil repellency, the dropped sample does not naturally flow due to capillary action and flows into the flow channel 5 by pressurization or suction operation. Here, the pressure operation and the suction operation method are arbitrary. For example, the pressure operation is realized by providing an elastic plug covering the sample supply port 4 and pressurizing it. This is realized by connecting a pump to a part of the second plate 2 for suction.

  After the sample reaches the measurement chamber 6, it undergoes a color reaction with the reagent held in the reagent holding unit 62. Here, the first plate 1 and the third plate 3 are configured to be transparent at the position of the measurement chamber 6. Accordingly, after the time required for completing the color reaction has elapsed, the measurement chamber is detected by detecting the incident light irradiated from the external light source toward the measurement chamber 6 and the transmitted light transmitted through the measurement chamber 6. The absorbance at 6 can be measured, and based on the absorbance obtained, the desired component in the sample can be quantified.

  Hereinafter, although it demonstrates using a more concrete Example, this invention is not limited to a following example.

Here, in order to confirm the influence of the difference in the materials used as the second plate 2 on the reproducibility of the absorbance, a test was conducted on the variation in absorbance when the following seven materials were used.
(1) Cellulose acetate membrane pore size 0.80 μm (manufactured by ADVANTEC, C080A)
(2) Cellulose acetate membrane, pore diameter 0.45 μm (ADVANTEC, C045A)
(3) Cellulose acetate membrane pore size 0.20 μm (manufactured by ADVANTEC, C020A)
(4) Cellulose mixed ester membrane, pore diameter 0.45 μm (ADVANTEC, A045A)
(5) Cellulose mixed ester membrane pore size 0.20 μm (A020A, manufactured by ADVANTEC)
(6) PTFE membrane (uniaxial stretching) Pore diameter 1.00 μm (Sumitomo Electric Fine Polymer, WP100)
(7) PTFE membrane (biaxial stretching) Pore diameter 1.00 μm (Sumitomo Electric Fine Polymer, FP100)
An analytical instrument is prepared using the above materials, and a blue dye solution (product name: Food Color Blue No. 1 (Brilliant Blue FCF, manufactured by Tokyo Chemical Industry Co., Ltd.), concentration: 0.04% (W / V) aqueous solution) is supplied as a sample. The sample was introduced into a measurement chamber, and the absorbance at a wavelength of 635 nm was measured. Since (1) to (5) are not inherently impervious to water, at least a substance mainly composed of a fluorocarbon group, a hydrocarbon group, and a chlorosilyl group, and a chlorosilyl group as a main component. Using a commercially available WR-LIFE1-3 (manufactured by Kagawa Student Venture Co., Ltd.), which is a composite film forming solution containing a substance or a substance mainly composed of an alkoxysilyl group and a non-aqueous organic solvent. Water and oil repellent treatment is applied. On the other hand, (6) and (7) are not subjected to water / oil repellent treatment because the material originally has water impermeability. The number of samples was 20 and the average absorbance, standard deviation, and coefficient of variation were determined. Table 1 shows the test results.

  As described above, the absorbance measurement using an analytical instrument employing a cellulose acetate membrane and a cellulose mixed ester membrane showed a small measurement coefficient and good measurement reproducibility. On the other hand, with respect to absorbance measurement using an analytical instrument employing a PTFE membrane, the coefficient of variation was large, and a large variation was clearly confirmed. This is due to the fact that the film thickness of the PTFE film itself varies greatly, and when the analytical instrument is created, the film thickness is likely to change due to processing such as lamination, and is difficult to restore. Therefore, it is preferable to use cellulose acetate or cellulose mixed ester as a porous material that is impermeable and breathable in order to realize a highly accurate analytical instrument with little variation in measurement results.

  Next, in order to confirm the change in the absorbance measurement value with the passage of time, the absorbance at a wavelength of 635 nm was measured for 10 minutes from the time when the sample was transferred to the measurement chamber 6 for the Example of the present invention and the Comparative Example. .

  These two differences are only in the water and oil repellent treatment of the second plate 2. In the embodiment of the present invention, the cellulose acetate membrane (pore size 0.80 μm, manufactured by ADVANTEC) is used as the second plate 2. WR-Live1-3 (manufactured by Kagawa Student Venture Co., Ltd.) was used for water / oil repellent treatment. On the other hand, in the comparative example, as the second plate 2, a fluororesin solution (FS-1010, Fluorotechnology), which is a general fluorine-based water and oil repellent treatment for a cellulose acetate membrane (pore size 0.80 μm, manufactured by ADVANTEC). Used).

  Parts other than the second plate 2 are the same, and the first plate 1 and the third plate 3 are screen-printed with carbon ink (SS Rio Phase C-A manufactured by Toyo Ink) on polyester film (Toray Lumirror) 188 μm (# 300 polyester, emulsion thickness: 15 μm), and a double-sided tape (5601 manufactured by Nitto Denko, base material PET: 4 μm, acrylic adhesive: 3 μm × 2 surfaces, total thickness 10 μm) was used for bonding the plate. . The length of the flow path 5 is 20 mm, and the width of the flow path 5 is 0.3 mm. The measurement chamber 6 has a diameter of 1.2 mm.

  As a sample, serum having a total cholesterol concentration of 220 mg / dL was introduced into the measurement chamber 6 by dropping 10 μL of the serum into the sample supply port 4 and applying pressure. When the sample reached the measurement chamber 6, the measurement of absorbance in light having a wavelength of 635 nm was started, and this measurement was continued for 10 minutes every 4 seconds to obtain a time course.

  FIG. 3 shows changes in absorbance measurement values with time. In the comparative example, since the sample is transferred to the measurement chamber 6 and the reagent starts to dissolve, the second plate 2 has been soaked, and the dye that has developed color moves to the plate. Is gradually decreasing. On the other hand, in the embodiment of the present invention, after the sample is transferred to the measurement chamber and the reagent is dissolved, the penetration into the second plate 2 does not occur, and the absorbance measurement value for 10 minutes is stable. Note that because of the surfactant contained in the reagent, the sample is not soaked in the second plate 2 even though the surface tension of the sample in which the reagent is dissolved is as small as 30 mN / m. It was confirmed that the oil function was sufficiently high.

Claims (8)

A sample supply port through which a sample is supplied;
A measurement chamber in which measurement of the sample is performed;
The flow path that connects the sample supply port and the measurement chamber,
The flow path and the measurement chamber are formed by stacking a plurality of plate members,
The plate forming at least the flow path among the plurality of plate members is a porous plate member having air permeability,
The surface of the porous plate member having air permeability includes at least a substance 1 mainly composed of a fluorocarbon group, a hydrocarbon group and a silyl group and a substance 2 mainly composed of a siloxane group on the surface. A body fluid component analyzer characterized by being covered with a chemically bonded water- and oil-repellent composite membrane. The body fluid component analysis instrument according to claim 1, wherein a material of the porous plate member having air permeability is cellulose acetate or cellulose mixed ester. The measurement chamber is provided with a reagent that reacts with a specific component in the body fluid and colors,
The analytical device for body fluid components according to claim 2, wherein the reagent contains a surfactant. A sample supply port through which a sample is supplied;
A measurement chamber in which measurement of the sample is performed;
The flow path that connects the sample supply port and the measurement chamber,
The flow path and the measurement chamber are formed by stacking a plurality of plate members,
In the method for producing a body fluid component analysis instrument, wherein the plate forming at least the flow path among the plurality of plate members is a porous plate member having air permeability,
The porous plate member having air permeability has at least a substance mainly composed of a fluorocarbon group, a hydrocarbon group and a chlorosilyl group, and a substance mainly composed of a chlorosilyl group or an alkoxysilyl group. A method for producing a body fluid component analysis instrument, comprising the step of contacting a composite film forming solution containing a substance and a non-aqueous organic solvent. A sample supply port through which a sample is supplied;
A measurement chamber in which measurement of the sample is performed;
The flow path that connects the sample supply port and the measurement chamber,
The flow path and the measurement chamber are formed by stacking a plurality of plate members,
In the method of manufacturing a body fluid component analysis instrument, the plate forming at least the flow path among the plurality of plate members is a porous plate member having air permeability.
The porous plate member having air permeability, a substance mainly containing at least a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group, a substance containing a silanol condensation catalyst or a chlorosilyl group, and a non-aqueous organic solvent A method for producing a body fluid component analytical instrument, comprising the step of bringing the composite film forming solution into contact. A sample supply port through which a sample is supplied;
A measurement chamber in which measurement of the sample is performed;
The flow path that connects the sample supply port and the measurement chamber,
The flow path and the measurement chamber are formed by stacking a plurality of plate members,
In the method of manufacturing a body fluid component analysis instrument, the plate forming at least the flow path among the plurality of plate members is a porous plate member having air permeability.
The porous plate member having air permeability is formed by using a substance mainly containing at least a fluorocarbon group, a hydrocarbon group, and an alkoxysilyl group, a substance mainly containing an alkoxysilyl group, a silanol condensation catalyst, or chlorosilyl. A method for producing a body fluid component analysis instrument, comprising a step of contacting a composite film forming solution containing a group-containing substance and a non-aqueous organic solvent. Obtained by the production method according to any one of claims 4 to 6,
The material of the porous plate member having air permeability is cellulose acetate or cellulose mixed ester,
The measurement chamber is provided with a reagent that reacts with a specific component in the body fluid and colors,
A surfactant is contained in the reagent. A body fluid component analysis instrument. Using the body fluid component analysis instrument according to claim 3 or 7,
Supplying a sample to the sample supply port;
Transferring the sample to the measurement chamber through the flow path;
And a step of measuring absorbance in the measurement chamber after the sample and the color sample have undergone a color reaction. A method for analyzing a body fluid component.

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