JP2003083926A - Apparatus and method for hemanalysis - Google Patents

Abstract

PROBLEM TO BE SOLVED: To solve the problem that although a small and portable blood autoanalyzer is used for a hemanalysis at present, a blood analyzer which sets a blood sampling amount required for the hemanalysis to be a trace amount in order to frequently perform the hemanalysis for the purpose of the daily health case of not only a patient but also a healthy person, and which reduces a pain and an inconvenience in an examination is required; that blood must be analyzed surely according to the blood sampled in the trace amount; that a small and satisfactory-sensitivity sensor corresponding to the blood in the trace amount is required; and that a calibration method suitable for the sensor is required in order to measure a component stably by the small sensor. SOLUTION: The sensor used for the hemanalysis is formed in such a way that an electrode is formed on the surface of an insulating-material substrate, that a part or the whole on the surface of the insulating-material substrate including the electrode is covered with a polymer membrane comprising a methyl group, and that a polymer membrane which contains ion-sensitive organic molecules or an enzyme is formed on the polymer membrane. A calibration solution used to calibrate the sensor is supplied from the outside of the blood analyzer, and the concentration of O2 and CO2 in the calibration solution is adjusted to the concentration of a normal value inside a body.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a blood analysis method and apparatus, and all of the functions and structures required for its operation are integrated in one device. Healthcare device characterized in that the device is small. In particular, the present invention relates to the structure and detection method of the detection means for detecting components in blood in the healthcare device, and the calibration method of the detection means.

[0002]

2. Description of the Related Art Thermometers, blood pressure monitors, ultrasonic diagnostics, X-ray CT, and
In addition to MRI, there is an automatic blood analyzer. this is,
Collect a few milliliters of blood and use a centrifuge to
Serum obtained by separating erythrocytes, leukocytes, lymphocytes, platelets, and blood coagulation factors is divided into a large number of test tubes, and the test tubes are moved in a line.
In addition to measuring each concentration of oxygen, carbon dioxide, etc., put reagents such as enzymes in the serum of each test tube and perform luminescence reaction absorption and absorption spectroscopy with the substrate in the serum and process the data with a computer It is used to diagnose.

The automatic blood analyzers are roughly classified into a flow system in which a sample is mixed with a reagent while passing through a tube and reacted to be conveyed to a detection unit, and a sample is dispensed into a container for each measurement item and supplied by a dispenser. There is a discrete method in which the reagent is mixed and reacted with the reagent to be quantitatively measured in the detection unit.
The mainstream of large-scale automatic analyzers is the discrete method, and the mainstream of tabletop and portable type analyzers is the flow method. The flow method has the advantages over the discrete method in that the device is simpler, easier to downsize, and requires less sample.

For example, in a large-scale blood analyzer used in a large-scale hospital or a blood test center facility, each biochemical test item has a volume of about 15 to 10 μl, and a desk-top type simple blood used in a practitioner or a small hospital. In the analyzer, it is about 10 to 5 μl per item. Recently,
It is the smallest portable type that has been used in the operating room and the bedside of hospitals.
μl.

The portable blood analyzer is 3 cm
A channel having a diameter of 1 mm is formed on two resin plates of about × 5 cm, and a cartridge containing a blood analysis sensor and a bag containing a calibration solution is provided in the middle of the channel, and a measuring instrument main body. The measurements are made in the following order. First, when about 50 μl of whole blood is injected into the cartridge and inserted into the main body, the calibration solution flows out from the calibration solution storage bag built in the cartridge to the analysis sensor section for calibration. The blood previously injected is introduced into the sensor unit in a state where it is separated by the calibration liquid and air bubbles, and measurement is performed. As a result, the values of electrolyte, glucose, O ₂ and CO ₂ in the blood are displayed on the display screen of the main body in about 2 minutes.

[0006] In this portable blood analyzer, the basic functions such as a calibration solution, a sensor, and an air storage tank for feeding blood / calibration solution are incorporated in a cartridge having a size of several cm to reduce a wasteful space through which blood passes. There are some ideas to reduce the amount of blood required, however, using this portable blood analyzer, not only the sick but also the healthy person can collect blood every day as part of daily health management and analyze the blood. However, it is difficult to say that the amount of blood required for analysis with this device is sufficiently small, and there has been a demand for the development of a blood analyzer capable of analyzing blood even from a smaller amount of blood.

Therefore, a small and simple blood analyzer (Japanese Patent Application No. 2000-120189) which has been developed in recent years uses an electroosmotic pump as a means for accurately moving a small amount of blood and a calibration solution. FIG. 1 shows an example thereof. Reference numeral 1 is a substrate, and 2 is a hollow needle for blood collection. Reference numeral 3 is a blood filtering means, and 4 is a separating means for obtaining serum from the filtered blood. Reference numeral 5 is a blood component analyzing means, and 6 is a flow path means. 7 is a moving means, and 8 and 9 are moving means 7 respectively.
It is an electrode insertion port for applying a voltage to.

[Problems to be Solved by the Invention]

In order to be able to analyze blood even from a small amount of blood, it is necessary to miniaturize the sensor used for blood analysis, and accordingly, the sensitive film used in the sensor also has a size of several tens of μm. A method of forming in degree is desired. In existing ion electrodes and enzyme sensors, the method of adhering the ion-sensitive film or enzyme film to the surface of the sensor substrate is mainly by using the chemical bonding force between the resist film and the ion-sensitive film or by increasing the surface area of the substrate and then the analytical sensor. The adhesion between the substrate and the ion-sensitive membrane or enzyme membrane is enhanced. However, there is no example in which a sensitive film having a thickness of several tens of μm is formed by such a conventional method, the sensor manufacturing process is often complicated, and it is necessary to find a simpler and more reliable adhesion method.

Further, in the blood analyzer shown in FIG. 1, although the movement controllability of the calibration solution used for the calibration of blood and the sensor is excellent, there is a problem in the method of introducing the calibration solution as an electrolytic solution into the moving means. Is left. In other words, if the direction from the blood collecting hollow needle to the analyzing means is set to the downstream side, the moving means is arranged on the downstream side of the analyzing means, so that when the calibration solution is introduced from the upstream side, the moving means and the blood collecting hollow needle are introduced. The air originally present in the flow path between the two enters the pump, causing a problem that the moving means does not operate.

Further, to calibrate the blood analysis sensor,
Sensors in dry storage must be immersed in the calibration solution. For this purpose, a nanometer-order calibrating solution is transported from the calibrating solution storage tank to the sensor section while controlling the position with an accuracy of about 10 μm, and the entire flow path having the sensor or a part thereof is filled with the calibrating solution. A liquid transfer method is required, the transfer means found in existing small blood analyzers cannot be used, and new means and control means for controlling this transfer means have been demanded.

Further, as a calibration liquid, a liquid in which a gas at a normal value level is dissolved is used for the calibration of a blood gas analysis sensor for O ₂ or CO ₂ , but in the unlikely event of a mistake in the operation procedure or malfunction of the device. The calibration solution must be safe even if it is injected into the body through the blood analyzer, and there is a problem to be solved in its preparation method.

[Means for Solving the Problems]

By forming a polymer film containing a methyl group on the surface of an insulating material substrate, an organic functional group of a polymer film containing an organic molecule or an enzyme which is ion-sensitive to the methyl group present on the outermost surface of the polymer film. The bond strength with the group enhances the adhesion of the film.

By providing a calibration solution storage tank and a pump for pumping the calibration solution outside the blood analyzer, it is possible to pump the calibration solution to the blood analyzer. Further, by introducing the calibration solution in the drain side of the electroosmotic flow pump, that is, in the direction from the pump toward the blood collecting needle, the calibration solution as the electrolytic solution can be filled in the electroosmotic flow pump without air bubbles being mixed therein. Also, by detecting the movement of this calibration solution,
It is possible to more reliably control the introduction of the calibration liquid.

Regarding the calibration solution, in order to calibrate the blood gas analysis sensor such as O ₂ or CO ₂ , the normal concentration of O ₂ and CO _{2 in} the body is dissolved in the solution serving as the calibration solution in advance, or By using O ₂ and CO ₂ gas when the calibration liquid is pressure-fed to the analytical sensor unit, the normal value concentration is dissolved in the liquid serving as the calibration liquid. In consideration of safety, the liquid is phosphate buffer (PBS) or physiological saline which is harmless to the human body and is used for treatment.

DETAILED DESCRIPTION OF THE INVENTION

FIG. 2 shows a schematic view of the analyzing means according to the present invention. Reference numeral 10 is an insulating material substrate, and an electrode 11 is formed on the insulating material substrate 10. Polymer film 1 having a methyl group on a part or all of the surface of insulating substrate 10 including electrode 11
Coat with 2. On the surface of the polymer film, a polymer film 13 containing an organic molecule or enzyme having ion sensitivity is formed.

FIG. 3 shows a schematic diagram of the blood analyzer according to the present invention. On the resin substrate 14, blood collecting means 15, moving means 16, separating means 17, analyzing means 18, flow path means 1
9. Form the filtering means 20. The calibration solution moving means 21 for pressure-feeding the calibration solution 22 for constituting the sensor constituting the analysis means 18 and the calibration solution accumulating means 23 for the calibration solution 22 are connected to the resin substrate 14 via the connecting means 24. Further, a gas supply means 25 for a gas containing O ₂ and CO ₂ is connected to the calibration liquid storage means 23. In order to collect the waste liquid of the calibration liquid used for the calibration of the sensor, a calibration liquid waste liquid recovery means 27 having a resin 26 having a water absorbing property is provided, and a connection means 28 is provided.
To connect with the sampling means 15. The flow path means 19 is provided with a plurality of means 29 for detecting the positions of the blood and the calibration solution. A valve 31 is provided in the calibration liquid pipe 30. A control means 32 for the calibration liquid and the blood flow rate is provided.

【Example】

[First Embodiment] In FIG. 2, an insulating substrate 1 is used.
After forming the electrode 11 on the 0 surface, a polymer film was formed using hexamethyldisilazane (HMDS) as the polymer film 12 having a methyl group on part or all of the substrate surface. Next, a solution in which a material for the organic film 13 containing an organic molecule or an enzyme having ion sensitivity is dissolved is applied and dried until the solvent evaporates. As a result, the peeling of the organic film from the insulating material substrate on which the electrode is formed is eliminated, and the instability of the output signal of the sensor constituting the analyzing means 18 due to the poor adhesion of the film is eliminated. Further, as the polymer film 12 having a methyl group, polyhydroxyethyl methacrylate (p-HEM) is used.
A polymer film was formed using A), but in this case as well, the instability of the output signal of the sensor constituting the analyzing means 18 disappeared.

[Second Embodiment] Analyzing means 1 in FIG.
8 is calibrated by the following procedure. The calibrating solution 22 contained in the calibrating solution accumulating means 23 is introduced into the flow path means 19 in the resin substrate 14 through the connecting means 24 by the moving means 21. This is a method of using a calibration liquid in which a gas containing O ₂ and CO ₂ is dissolved in advance when calibrating the analysis means 18, and the calibration liquid 22 is normally fed while being pumped with the gas containing O ₂ and CO _2. In the method of dissolving the gas at the value level, the calibration liquid 22 is generated by the gas containing O ₂ and CO ₂ in the gas supply means 25.
May be pumped to the flow path means 19. As the calibration solution 22, a phosphate buffer solution (PB) that is harmless to the human body and is used for treatment is used.
S) or using physiological saline is used. Then, when the calibration solution 22 reaches the detecting means 29 provided in the flow path means 19, the driving of the moving means 21 is stopped. Alternatively, when introducing the calibration liquid using the moving means 16, the supply of the gas containing O ₂ and CO ₂ is stopped. Detection means 29 used here
May be a method of detecting a change in the output signal of the analyzing means 18, an optical method of combining an LED light emitting element and a light receiving element, or a method of detecting an electric capacity. Then, the analysis means 18 is calibrated. When the calibration is completed, the moving means 16 causes the calibration solution 22 in the flow path means 19 to pass through the connecting means 28 and the resin 26 in the calibration solution waste solution collecting means 27.
Absorb and collect. This prevents the calibration liquid from flowing back and the calibration liquid from flowing out and becoming dirty around the device. In addition, the calibration liquid 22 is collected by the gas supply means 25.
The calibration liquid 22 may be discharged from the inside of the apparatus by the pressure of the gas supplied from the device.

[Third Embodiment] FIG. 4 is a modification of the second embodiment and has a structure in which the calibration liquid waste liquid recovery means 27 shown in FIG. 3 is connected to the gas supply means 25. 33 is a resin substrate, 34 is a separating means, 35 is a blood collecting hollow needle, 36 is a flow path means, and 37 is a moving means. 38 is a connecting means, 39 is a valve, 40 is a calibration solution, 41 is a calibration solution accumulating means, 42 is a gas supply means, 43 is a calibration solution waste solution collecting means, 44 is a water-absorbing resin, and 45 is an analyzing means. Analyzing means 45
The calibration solution is supplied to the second embodiment in the same manner. That is, the calibration liquid 40 that has entered the calibration liquid storage means 41.
Is introduced into the flow path means 36 in the resin substrate 33 through the connection means 38 by the gas supply means 42. Further, the calibration liquid 40 is supplied by the gas containing O ₂ and CO ₂ in the gas supply means 42.
May be pumped to the flow path means 36. After the calibration of the detection means 45 provided in the flow path means 36, the used calibration liquid 40 is moved to the calibration liquid waste liquid collection means 43 by the movement means 37 and collected. Further, the used calibration liquid 40 may be sucked using the pump 46 and moved to the calibration liquid waste liquid recovery means 43 to be recovered. As a result, as in the second embodiment, the calibration liquid 40 does not flow back and the calibration liquid does not flow out around the device and become dirty.

[0019]

According to the present invention, the sensitive film of the microanalytical sensor can be manufactured with good reproducibility, and the analytical sensor can be calibrated in the micro blood analyzer.

[Brief description of drawings]

FIG. 1 is a diagram for explaining a conventional device.

FIG. 2 is a diagram illustrating a structure for improving the adhesion of an organic film containing an organic molecule or an enzyme having ion sensitivity to an insulating material substrate.

FIG. 3 is a schematic diagram of an apparatus according to the present invention.

FIG. 4 is a diagram for explaining an embodiment of the device according to the present invention.

[Explanation of symbols]

DESCRIPTION OF SYMBOLS 1 Substrate 2 Hollow needle for blood collection 3 Filtration means 4 Separation means 5 Analysis means 6 Flow path means 7 Moving means 8 Electrode insertion port 9 Electrode insertion port 10 Insulating material substrate 11 Electrode 12 Polymer film 13 having methyl group 13 Ion sensitivity Polymer film containing organic molecule or enzyme 14 Resin substrate 15 Collection means 16 Transfer means 17 Separation means 18 Analysis means 19 Flow path means 20 Filtration means 21 Calibration solution transfer means 22 Calibration solution 23 Calibration solution storage means 24 Connection means 25 Gas supply means 26 Resin 27 Calibration liquid waste liquid collection means 28 Connection means 29 Detection means 30 Calibration liquid piping 31 Valve 32 Control means 33 Resin substrate 34 Separation means 35 Hollow needle 36 Flow path means 37 Moving means 38 Connection means 39 Valve 40 Calibration Liquid 41 Calibration liquid storage means 42 Gas supply means 43 Calibration liquid waste liquid collection means 44 Resin 45 Analysis means 46 Pump

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) G01N 33/483 G01N 27/30 353A 353J (72) Inventor Madoka Takai 2-18 Minamiotsuka, Toshima-ku, Tokyo 24-305 (72) Inventor Zen Takamura Zen, 4-9-9 Minamisenju, Arakawa-ku, Tokyo River Harp Minamisenju 401 (72) Inventor Hiroshi Otsuka 2-5-51 Aihara, Sagamihara-shi, Kanagawa (72) Inventor Ogawa Hiroki 2-18, Shin-Yokohama, Kohoku-ku, Yokohama-shi, Kanagawa 1 Century Shin-Yokohama Room 701 (72) Inventor Jun Kikuchi 2-14-F, Shirokanedai, Minato-ku, Tokyo 2F0 (Reference) 2G045 AA01 BA08 CA25 DB30 FB05 JA02 JA07

Claims (12)

[Claims] 1. A collection means for collecting blood from a living body, a filtration means for filtering at least the collected blood to obtain plasma, or a separation means for separating serum from the blood in one or a plurality of substrates. Any means, an analyzing means for analyzing a substance in the blood, the collecting means, the filtering means, the separating means, a flow path means for connecting the analyzing means, the collecting means, the filtering means, the Separation means, the analysis means, moving means for moving the components of the blood present in the flow path means, output means for taking out the information from the analysis means to the outside, the collection means, the filtration means, the separation A control means for controlling the operation of at least one of the means, the analyzing means, the moving means and the output means, and a holding means for holding the blood component in the substrate, and In the case of a plurality of substrates, in the blood analyzer characterized by having a structure in which the substrates are integrated, the analysis means, the electrodes on the insulating substrate via the organic molecular film having a methyl group A blood analyzer characterized by being formed with a polymer film containing an organic molecule or an enzyme having ion sensitivity. 2. A collection means for collecting blood from the inside of a living body, a filtration means for filtering at least the collected blood to obtain plasma, or a separation means for separating serum from the blood in one or a plurality of substrates. Any means, an analyzing means for analyzing a substance in the blood, the collecting means, the filtering means, the separating means, a flow path means for connecting the analyzing means, the collecting means, the filtering means, the Separation means, the analysis means, moving means for moving the components of the blood present in the flow path means, output means for taking out the information from the analysis means to the outside, the collection means, the filtration means, the separation A control means for controlling the operation of at least one of the means, the analyzing means, the moving means and the output means, and a holding means for holding the blood component in the substrate, and In the case of a plurality of substrates, in a blood analysis method characterized in that blood is collected, filtered, separated, and analyzed by a blood analyzer having a structure in which the substrates are integrated, an insulating material is used as the analysis means. A blood analysis method, characterized in that blood is analyzed by an analysis means in which a polymer film containing an organic molecule or an enzyme having ion sensitivity is formed on an electrode on a substrate through an organic molecule film having a methyl group. 3. In one or a plurality of substrates, a collecting means for collecting blood from the inside of a living body, a filtering means for filtering at least the collected blood to obtain plasma, or a separating means for separating serum from the blood. Any means, an analyzing means for analyzing a substance in the blood, the collecting means, the filtering means, the separating means, a flow path means for connecting the analyzing means, the collecting means, the filtering means, the Separation means, the analysis means, moving means for moving the components of the blood present in the flow path means, output means for taking out the information from the analysis means to the outside, the collection means, the filtration means, the separation A control means for controlling the operation of at least one of the means, the analyzing means, the moving means and the output means, and a holding means for holding the blood component in the substrate, and In a case where there are a plurality of substrates, the blood analyzer has a structure in which the substrates are integrated, in the blood analysis device, the analysis means first forms an electrode on an insulating material substrate, and then the insulating material substrate. Manufacture of a blood analyzer characterized by forming an organic molecular film having a methyl group on at least the surface of an electrode, and then forming a polymer film containing an organic molecule or an enzyme having ion sensitivity Method. 4. The organic molecule film having a methyl group according to claim 1, wherein hexamethyldisilazane (HMDS) or polyhydroxyethyl methacrylate (p-HEM) is used.
A blood analyzer characterized by being an organic molecular film according to A). 5. The blood analysis method according to claim 2, wherein a calibration solution is introduced from the outside of the blood analyzer to calibrate the analysis means. 6. A blood analysis method, wherein the calibration liquid according to claim 5 is pressure-fed by the moving means from the downstream side of the flow path means of the blood analyzer. 7. A blood analysis method, wherein the calibration liquid according to claim 5 is a liquid in which a gas containing O ₂ and CO ₂ molecules having a concentration close to that in blood in a living body is dissolved in advance. 8. The calibration solution according to claim 5 contains O ₂ and C.
A blood analysis method, wherein a gas containing O ₂ molecules is injected under pressure, and O ₂ and CO ₂ having normal concentration in the body are dissolved. 9. A blood analysis method, wherein the calibration solution according to claim 5 is a phosphate buffer solution (PBS) or physiological saline. 10. The blood analyzer according to claim 1, further comprising a calibration liquid waste liquid recovery means for recovering the calibration liquid waste liquid used for the calibration of the analysis means, outside the blood analysis device. Blood analyzer. 11. A blood analyzer, wherein the calibration liquid waste liquid collecting means according to claim 10 comprises a resin having a water absorbing property. 12. A collection means for collecting blood from within a living body, a filtration means for filtering at least the collected blood to obtain plasma, or a separation means for separating serum from the blood in one or a plurality of substrates. Any means, an analyzing means for analyzing a substance in the blood, the collecting means, the filtering means, the separating means, a flow path means for connecting the analyzing means, the collecting means, the filtering means, the Separation means, the analysis means, moving means for moving the components of the blood present in the flow path means, output means for taking out the information from the analysis means to the outside, the collection means, the filtration means, the separation A control means for controlling the operation of at least one of the means, the analysis means, the moving means, and the output means, and a holding means for holding the components of the blood in the substrate. In the case of a plurality of substrates, the blood analyzer has a structure in which the substrates are integrated, and a plurality of detecting means for detecting the presence or absence of a liquid inside the flow passage means of the analyzing means. A blood analyzer characterized in that the moving means is controlled in accordance with a detection signal from the detecting means.