WO2012133394A1 - 試料溶液濃度測定方法及び試料溶液濃度測定装置 - Google Patents
試料溶液濃度測定方法及び試料溶液濃度測定装置 Download PDFInfo
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- WO2012133394A1 WO2012133394A1 PCT/JP2012/057913 JP2012057913W WO2012133394A1 WO 2012133394 A1 WO2012133394 A1 WO 2012133394A1 JP 2012057913 W JP2012057913 W JP 2012057913W WO 2012133394 A1 WO2012133394 A1 WO 2012133394A1
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/62—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving urea
Definitions
- the present invention relates to a sample solution concentration measuring method for quantifying the concentration of a sample solution by measuring chemiluminescence generated by mixing the sample solution and the reactant solution, and a sample solution concentration measuring apparatus used therefor.
- a tubular hollow fiber assembly called a dialyzer made of a semipermeable membrane having an inner diameter of about 100 ⁇ m is used. Solutes such as urea present in the blood are removed from the body by permeating into the dialysate side while passing through the dialyzer. Conventionally, the amount of urea in blood has been analyzed by collecting blood, and has been evaluated as a blood urea nitrogen (hereinafter sometimes referred to as “BUN”) value.
- BUN blood urea nitrogen
- a colorimetric method that measures the urea concentration by using a reagent that develops color by reacting with urea and comparing it with the standard color of the reagent, specific to urea
- An enzyme thermistor method that measures urea concentration by immobilizing urease, which is a working enzyme, around glass beads and measuring the heat of reaction accompanying the hydrolysis reaction of urea, an oxidizing agent that reacts with urea to produce chemiluminescence, for example
- a chemiluminescence method for measuring urea concentration based on the intensity of chemiluminescence (hereinafter sometimes abbreviated as “CL”) using hypohalite is known.
- the enzyme thermistor method the enzyme deteriorates every time the measurement is repeated, the change of the enzyme over time is large, and it is difficult to measure stably for a long period of time. Although it can be used for monitoring the urea concentration, there is a problem in accuracy when used for multiple measurements.
- Non-Patent Document 1 Chemiluminescence is said to occur when excited nitrogen produced during the reaction of urea and hypohalite ions returns to the ground state.
- Non-Patent Document 2 describes measuring the urea concentration in the dialysis waste liquid as a guide for knowing the BUN value.
- Non-Patent Document 2 describes a method for measuring urea concentration by measuring chemiluminescence generated by the reaction of urea in dialysis waste liquid with sodium hypobromite.
- chemiluminescence efficiency and reproducibility are not always good, and improvements have been demanded.
- a chemiluminescence measuring method (flow injection method) using a reaction vessel in which a glass tube is processed into a spiral shape is known.
- the sample solution and the reactant solution are sent to the introduction port of the reaction vessel by a pump or the like, and the sample solution and the reactant solution are mixed and introduced into the spiral tube at the introduction port, and the generated chemiluminescence is measured. Is the method.
- the sample solution and the reactant solution cannot be mixed uniformly in a short time. Especially when the sample solution and the reactant solution are mixed, a reaction with a short chemiluminescence lifetime occurs or bubbles are generated. When such a reaction occurs, the reproducibility of the measurement of the chemiluminescence intensity is not good and an improvement has been demanded.
- Patent Document 1 discloses a chemiluminescence measurement method for measuring chemiluminescence generated in a cylinder, in which one of a sample solution and a reactant solution is sucked into the cylinder by moving a piston in the cylinder, and then Chemiluminescence characterized by inhaling the other solution and generating a turbulent flow in the cylinder by the jet of the other solution to uniformly mix the sample solution and the reactant solution.
- the measurement method is described. According to this method, chemiluminescence measurement with high luminous efficiency and good reproducibility can be performed.
- Patent Document 2 includes a plurality of independent reaction chambers formed by anisotropic etching on the surface of a silicon substrate, and a flat plate that is anodically bonded to the surface of the silicon substrate and seals the reaction chamber.
- the bioreactor microreactor is characterized by the following: According to this, for example, a large number of biochemical reactions of 1000 or more can be performed simultaneously in parallel, and not only simple analysis but also substance synthesis reactions such as protein synthesis can be performed on the cell. ing. In addition, it is said that biochemical reactions can be optically observed and the emission intensity can be monitored. However, in the above microreactor aiming at finally obtaining a reaction product, there are cases where uniform mixing cannot be performed in a short time in each independent reaction chamber.
- the present invention has been made to solve the above-mentioned problems, and it is possible to accurately determine the concentration of a sample solution by measuring the chemiluminescence intensity generated by mixing the sample solution and the reagent solution with good reproducibility. It aims at providing the measuring method which can be performed. Moreover, it aims at providing the suitable use of such a measuring method.
- An object of the present invention is to provide a sample solution concentration measurement method for quantifying the concentration of a sample solution by measuring the chemiluminescence intensity generated in the reaction vessel, wherein the reaction vessel has 2 reaction tanks each having an ejection hole and a discharge hole. Two or more are connected in series, and the cross-sectional area ratio (S1 / S2) between the cross-sectional area (S1) of each of the reaction vessels and the cross-sectional area (S2) of the ejection hole is 3 or more, The solution and the reactant solution are introduced into the first reaction tank from the ejection hole of the first reaction tank, and a turbulent flow is generated in the first reaction tank by the jet of the introduced solution to react the sample solution with the sample solution.
- the first mixed solution obtained is mixed into the second reaction tank through the discharge hole of the second reaction tank through the discharge hole of the first reaction tank, and the first mixed solution is mixed with the agent solution.
- a turbulent flow is generated in the second reaction tank by the jet of the first mixed solution
- the reaction vessel optionally has a configuration in which one or more additional reaction vessels are repeatedly arranged downstream of the second reaction vessel, the above in the second reaction vessel
- the mixed solution may be further mixed in the same manner as the mixing, and this is solved by providing a sample solution concentration measuring method characterized by measuring the chemiluminescence intensity generated in the reaction vessel.
- the sample solution is preferably a sample solution containing urea, and the sample solution is preferably a dialysis waste liquid. It is also preferable that the reactant solution is a reactant solution containing hypohalite ions.
- the above-described problem is a sample solution concentration measuring apparatus for quantifying the concentration of a sample solution by measuring the intensity of chemiluminescence generated in a reaction vessel, wherein two or more reaction tanks having ejection holes and discharge holes are connected in series.
- a reaction vessel connected to each other, the cross-sectional area ratio (S1 / S2) between the cross-sectional area (S1) of each of the reaction vessels and the cross-sectional area (S2) of the ejection holes is 3 or more, and the reaction vessel
- a turbulent flow is generated in each of the reaction tanks by a jet of the introduced solution, and the chemiluminescence intensity generated in the reaction vessel is measured by mixing the sample solution and the reactant solution.
- Sample solution concentration measuring device Also solved by providing.
- An artificial dialysis apparatus that quantifies the urea concentration in the dialysis waste liquid by the sample solution concentration measuring apparatus is a preferred embodiment of the present invention.
- the mixing of the sample solution and the reactant solution is repeated by the jet of the solution introduced into each reaction tank.
- the chemiluminescence intensity generated by the mixing can be measured with high reproducibility, the concentration of the sample solution can be accurately quantified.
- the reaction rate of chemiluminescence is fast, the chemiluminescence intensity can be measured with good reproducibility.
- the chemiluminescence intensity can be measured with good reproducibility even when bubbles are generated along with the chemiluminescence reaction when the sample solution and the reactant solution are mixed.
- the sample solution is a sample solution containing urea
- the urea concentration can be measured in real time, so that it can be suitably used as an artificial dialysis apparatus that can know the end of the dialysis time.
- FIG. 2 is a chemiluminescence response waveform diagram obtained in Example 1.
- FIG. 3 is a chemiluminescence response waveform diagram obtained when the concentration of an aqueous urea solution was changed in Example 1.
- FIG. 2 is a chemiluminescence response waveform diagram used for calculation of chemiluminescence lifetime ⁇ in Example 1.
- FIG. 3 is a correlation diagram between the urea concentration obtained in Example 1 and the chemiluminescence intensity.
- 6 is a chemiluminescence response waveform diagram obtained in Example 2.
- FIG. 6 is a chemiluminescence response waveform diagram obtained in Comparative Example 1.
- FIG. FIG. 2 is a schematic plan view of an apparatus for measuring chemiluminescence intensity generated in each reaction vessel A to G.
- FIG. 6 is a chemiluminescence response waveform diagram obtained by measuring the chemiluminescence intensity generated in each reaction vessel A to G.
- FIG. 1 is a schematic plan view showing an example of a sample solution concentration measuring apparatus 1 used in the present invention
- FIG. 2 is a schematic side view showing an example of the apparatus 1.
- a reaction vessel 2 is provided in which reaction vessels having ejection holes and discharge holes from the first reaction vessel A to the seventh reaction vessel G are connected in series.
- Each reaction vessel has a transparent portion on the upper surface side of the reaction vessel 2, and a photodetector 24 is arranged at a position facing the upper surface of the reaction vessel 2 as shown in FIG. 2.
- the sample solution inlet 3 and the first reaction tank A are connected by a pipe 7.
- the sample solution is introduced into the first reaction tank A from the ejection hole 9 of the first reaction tank A through the pipe 7 from the sample solution introduction port 3 by operating the pump 5.
- the reactant solution inlet 4 and the first reaction tank A are connected by a pipe 8.
- the reactant solution is introduced into the first reaction tank A from the ejection hole 9 of the first reaction tank A through the pipe 8 from the reactant solution introduction port 4 by operating the pump 6.
- a turbulent flow is generated in the first reaction tank A by the jet of the solution introduced into the first reaction tank A, and the sample solution and the reactant solution are uniformly mixed, so that the first mixed solution becomes can get.
- the obtained first mixed solution is introduced into the second reaction tank B from the ejection hole 11 of the second reaction tank B through the discharge hole 10 of the first reaction tank A.
- a turbulent flow is generated in the second reaction tank B by the jet flow of the introduced first mixed solution, and the first mixed solution is further mixed to obtain a second mixed solution.
- the obtained second mixed solution is contained in each reaction tank from the third reaction tank C to the seventh reaction tank G provided downstream of the discharge hole 12 of the second reaction tank B. Further mixed.
- the chemiluminescence intensity generated by mixing the sample solution and the reagent solution in the reaction vessel 2 is measured by the photodetector 24, and the concentration of the sample solution can be quantified.
- the sample solution concentration measuring apparatus 1 of the present invention includes a reaction vessel 2 in which two or more reaction vessels having ejection holes and discharge holes are connected in series.
- a reaction vessel 2 in which two or more reaction vessels having ejection holes and discharge holes are connected in series.
- the reaction vessel 2 may optionally have a configuration in which one or more additional reaction vessels are repeatedly arranged downstream of the second reaction vessel B.
- the reproducibility of the measurement of the chemiluminescence intensity is improved by repeatedly arranging the plurality of reaction vessels.
- the reaction vessel 2 used in the present invention it is preferable that three or more of the reaction vessels are connected in series, more preferably four or more are connected in series, and more than five are connected in series. More preferably, it is connected to.
- the concentration of the sample solution is low, the reproducibility of the measurement of chemiluminescence intensity is better when the number of connected reaction vessels is larger.
- the shape of the reaction vessel used in the present invention is not particularly limited.
- the shape of the reaction vessel may be a cylindrical shape, a spherical shape, or a square shape. From the viewpoint of good stirring efficiency, a cylindrical or spherical reaction vessel is preferably used.
- the size of the reaction vessel used in the present invention is not particularly limited, but from the viewpoint of efficiently generating turbulent flow by the jet of the solution and reducing the amount of the reactant solution used, the capacity of one reaction vessel Is preferably 300 mm 3 or less, more preferably 200 mm 3 or less, and even more preferably 100 mm 3 or less.
- the sample solution concentration measuring apparatus 1 of the present invention preferably has at least one reaction vessel in which an angle ⁇ between the axis of the ejection hole and the axis of the discharge hole is 15 degrees or more and 165 degrees or less.
- the cross-sectional area ratio (S1 / S2) between the cross-sectional area (S1) of each reaction vessel and the cross-sectional area (S2) of the ejection hole is 3 or more.
- the cross-sectional area (S1) of the reaction tank means the maximum cross-sectional area in a plane perpendicular to the water flow flowing into the reaction tank from the ejection holes.
- the sample solution and the reactant solution are rapidly mixed and homogenized, so that fluctuations in the chemiluminescence intensity when measuring the generated chemiluminescence intensity are reduced, the S / N ratio is improved, and the chemiluminescence is increased.
- the reproducibility of intensity measurement is good.
- the cross-sectional area ratio (S1 / S2) is less than 3, turbulent flow and laminar flow are mixed when the sample solution and the reactant solution are introduced into the reaction vessel from the ejection holes, and the chemiluminescence intensity is measured.
- the reproducibility may be reduced, and the cross-sectional area ratio (S1 / S2) is preferably 4 or more, and more preferably 5 or more.
- the cross-sectional area ratio (S1 / S2) is usually 1000 or less.
- each reaction tank has a transparent portion, and a photodetector 24 is disposed outside the transparent portion.
- the light detector 24 measures the chemiluminescence generated in the reaction vessel 2 by mixing the sample solution and the reactant solution.
- concentration of the sample solution can be quantified by measuring the chemiluminescence intensity using the photodetector 24.
- the arrangement of the light detector 24 is not particularly limited as long as it is outside the transparent portion of each reaction tank. However, as shown in FIG. 2, the light detector 24 is arranged so as to be in contact with the upper surface of the reaction vessel. It is preferable to do.
- the material constituting the reaction vessel used in the present invention is not particularly limited.
- the material constituting the reaction vessel may be glass, acrylic resin, polystyrene resin, polyvinyl chloride resin, polycarbonate resin, polyester. It may be a transparent resin such as a resin, or a combination of these.
- a thermoplastic resin is preferably used from the viewpoint of easy molding and low cost.
- the sample solution concentration measuring apparatus 1 of the present invention by using the sample solution concentration measuring apparatus 1 of the present invention for a long period of time, even if contaminants accumulate inside the reaction tank, it can be manufactured at low cost and can be used disposable.
- the material constituting the reaction vessel is a thermoplastic resin such as an acrylic resin
- the present inventors once distribute the strongly alkaline reactant solution into the reaction vessel, and then the sample solution and the reactant.
- the chemiluminescence intensity generated by mixing with the solution was measured, it was confirmed that the reproducibility of the obtained chemiluminescence intensity was improved. The reason for this is not necessarily clear, but the present inventors speculate that there is a possibility that the surface in the reaction vessel may be hydrophilized by the reaction solution having strong alkalinity.
- the sample solution concentration measuring apparatus 1 of the present invention includes means for introducing the sample solution and the reactant solution into the first reaction tank A from the ejection holes 9.
- the introduction means is not particularly limited as long as the sample solution and the reactant solution can be introduced into the first reaction tank A at a constant flow rate.
- a diaphragm pump, a piston pump, a plunger pump, a gear pump, etc. are mentioned.
- a diaphragm pump is preferably used from the viewpoint of good responsiveness and quantitativeness. Since the sample solution concentration measuring apparatus 1 of the present invention does not require any mechanical operation means other than the introduction means, the apparatus configuration is not complicated and the concentration of the sample solution can be determined without maintenance. . In addition, the sample solution concentration measuring apparatus 1 of the present invention can be manufactured at a low cost and hardly break down.
- the sample solution concentration measuring apparatus 1 of the present invention may include a sample solution supply unit capable of supplying a sample solution, for example, a tank in which the sample solution is stored. From the viewpoint of quantifying the concentration of the sample solution in real time, it is preferable that the sample solution supply unit includes means capable of supplying a new sample solution as needed. As will be described later, when the sample solution is a sample solution containing urea, particularly a dialysis waste liquid, a new dialysis waste liquid can be supplied at any time, so that the urea concentration in the dialysis waste liquid can be quantified in real time. This makes it possible to know the timing at which dialysis treatment should be terminated. Moreover, the sample solution concentration measuring apparatus 1 of the present invention may include a reactant solution supply unit capable of supplying a reactant solution, and includes, for example, a tank in which the reactant solution is accommodated.
- the concentration of the sample solution is quantified by measuring the intensity of chemiluminescence generated in a reaction vessel 2 in which two or more reaction vessels having ejection holes and discharge holes are connected in series. It is characterized by doing.
- the turbulent flow is generated in each reaction vessel by the jet of the solution introduced into the reaction vessel, so that the sample solution and the reactant solution are rapidly mixed and uniformed.
- the fluctuation of the chemiluminescence intensity when measuring the chemiluminescence intensity is reduced, the S / N ratio is improved, and the reproducibility of the chemiluminescence intensity measurement is improved.
- the sample solution concentration can be quantified.
- the sample solution and the reactant solution are introduced into the first reaction tank A, and the first reaction tank is jetted by the introduced solution.
- a turbulent flow is generated in A, and the sample solution and the reactant solution are uniformly mixed to obtain a first mixed solution.
- the obtained first mixed solution is introduced into the second reaction tank B from the ejection hole 11 through the discharge hole 10.
- a turbulent flow is generated in the second reaction tank B by the jet flow of the introduced first mixed solution, and the first mixed solution is further mixed to obtain a second mixed solution.
- the obtained second mixed solution is further mixed in each reaction tank from the third reaction tank C to the seventh reaction tank G provided downstream of the discharge hole 12. Then, the mixed solution is discharged from the discharge port 23.
- the concentration of the sample solution can be quantified by measuring the chemiluminescence intensity generated by mixing the sample solution and the reactant solution in the reaction vessel 2 with the photodetector 24.
- the chemiluminescence intensity generated by the mixing can be measured with good reproducibility.
- the reproducibility of the measurement of the chemiluminescence intensity is good even when bubbles are generated along with the reaction that generates chemiluminescence by mixing the sample solution and the reactant solution.
- the present inventors have confirmed that when bubbles are generated when the sample solution and the reactant solution are mixed, the stability of chemiluminescence intensity was not good, and the reaction was inhibited by the generated bubbles. It is presumed that the chemiluminescence is scattered.
- the present inventors have shown that bubbles are generated when the sample solution and the reagent solution are mixed in the first reaction tank A to obtain the first mixed solution. It is confirmed visually. When the bubbles generated together with the first mixed solution pass through the respective reaction tanks from the second reaction tank B to the seventh reaction tank G, it is difficult to visually confirm the generated bubbles. It was. This is considered to be because the bubbles generated with the first mixed solution were refined by repeating the mixing by the jet of the solution in each reaction tank. On the other hand, as can be seen from the results of Comparative Example 1 described later, when a reaction vessel having only one reaction vessel is used, the stability of the chemiluminescence intensity generated by mixing the sample solution and the reactant solution is not good. There wasn't. Therefore, it is a preferred embodiment of the present invention to quantify the concentration of the sample solution by measuring the chemiluminescence intensity generated in the reaction vessel 2 when the reaction that generates bubbles occurs.
- the sample solution concentration measurement method of the present invention has good reproducibility of chemiluminescence intensity measurement even when the reaction rate of chemiluminescence generated by mixing the sample solution and the reagent solution is high. Therefore, the concentration of such a sample solution can be quantified with high accuracy.
- a fast reaction rate of chemiluminescence indicates that the chemiluminescence lifetime ⁇ is small.
- the chemiluminescence lifetime ⁇ is the chemiluminescence when the decay of the chemiluminescence intensity occurs. It is defined as the time until the intensity decreases to a value of 1 / e. As can be seen from the chemiluminescence response waveform diagram of FIG.
- the chemiluminescence intensity generated by mixing the sample solution and the reactant solution attenuates exponentially when the supply of the sample solution and the reactant solution is stopped.
- the chemiluminescence lifetime ⁇ is obtained until the chemiluminescence intensity on the decay curve decreases to a value of 1 / e in the chemiluminescence response waveform diagram in which the vertical axis in the semilogarithmic graph is chemiluminescence intensity and the horizontal axis is time. It is calculated by the time.
- the reason why the reproducibility of the measurement of chemiluminescence intensity is good is that turbulent flow is generated in each reaction tank by a jet of solution introduced into the reaction tank.
- a preferred embodiment of the present invention is a sample solution concentration measurement method in which the concentration of a sample solution is quantified by measuring the intensity of chemiluminescence generated in the reaction vessel 2 in a chemiluminescence reaction having a chemiluminescence lifetime ⁇ of 3 seconds or less. It is an aspect.
- the sample solution used in the present invention is not particularly limited as long as it is chemiluminescent by mixing with a reactant solution, but it may be a sample solution containing a biologically derived nitrogen-containing compound such as urea, amino acid, protein, etc. A sample solution containing urea is more preferable.
- the sample solution is a sample solution containing urea
- the sample solution concentration measuring device 1 of the present invention can be suitably used for various applications as a urea concentration measuring device.
- the reagent solution used in the present invention is not particularly limited as long as it is chemiluminescent by mixing with the sample solution, but it reacts with the sample solution containing a biologically derived nitrogen-containing compound such as urea, amino acid, protein and the like.
- the reactant solution is preferably a reactant solution containing hypohalite ions.
- the hypohalite ion is not particularly limited, and examples thereof include hypohalite ions such as FO ⁇ , ClO ⁇ , BrO ⁇ , IO ⁇ , etc., and are selected from hypobromite ions or hypochlorite ions. It is preferable that there is at least one.
- the reactant solution containing hypohalite ions may be supplied to the apparatus in advance as an aqueous solution containing hypohalite ions, or the aqueous solution containing halogen ions is electrolyzed in the apparatus. You may supply by.
- the sample solution used in the present invention is a dialysis waste liquid
- it can be suitably used as an artificial dialysis apparatus characterized by measuring the urea concentration in the dialysis waste liquid.
- the urea concentration can be measured in real time
- it can be suitably used as an artificial dialysis apparatus that can know the timing at which dialysis treatment should be terminated.
- the method for evaluating the performance of a dialyzer using the sample solution concentration measuring apparatus 1 of the present invention is also a preferred embodiment of the present invention.
- FIG. 1 is a schematic plan view showing an example of a sample solution concentration measuring apparatus 1
- FIG. 2 is a schematic side view of the apparatus 1.
- the reaction vessel 2 (vertical: 40 mm, horizontal: 40 mm, height: 5 mm) has a cylindrical reaction vessel (diameter) from the first reaction vessel A to the seventh reaction vessel G. : 3 mm, height: 3 mm) are connected in series. Glass plates were placed on the upper and lower surfaces of the acrylic resin plate on which the reaction vessels from the first reaction vessel A to the seventh reaction vessel G were formed.
- each reaction tank from the first reaction tank A to the seventh reaction tank G is provided with an ejection hole (inner diameter: 1 mm) and a discharge hole (inner diameter: 1 mm).
- the cross-sectional area ratio (S1 / S2) between the cross-sectional area (S1) of each reaction tank and the cross-sectional area (S2) of the ejection hole was 9.
- the discharge hole 10 of the first reaction tank A and the ejection hole 11 of the second reaction tank B are connected via a narrow passage, and a third reaction is further provided downstream of the discharge hole 12 of the second reaction tank B.
- Respective reaction tanks from the tank C to the seventh reaction tank G are connected in series.
- the angle ⁇ in each reaction tank is 90 degrees in the reaction tank A, 120 degrees in the reaction tank B, 60 degrees in the reaction tank C, 180 degrees in the reaction tank D, 60 degrees in the reaction tank E, and 60 degrees in the reaction tank F.
- the reaction tank G was 120 degrees and 120 degrees.
- a photosensor module which is a photodetector 24, was attached at a position facing the upper surface of the reaction vessel 2 so as to be in contact with the upper surface.
- the sample solution inlet 3 is connected to the first reaction tank A via a pipe 7 (inner diameter: 1 mm), and the reactant solution inlet 4 is connected to the first reaction tank A via a pipe 8 (inner diameter: 1 mm). It is connected.
- the sample solution is introduced into the first reaction tank A from the sample solution introduction port 3 through the pipe 7 by operating the pump 5.
- the reactant solution is introduced into the first reaction tank A from the reactant solution inlet 4 through the pipe 8 by operating the pump 6.
- FIG. 1 the configuration in which the pipe 7 and the pipe 8 merge immediately before the sample solution and the reactant solution are introduced into the first reaction tank A from the ejection holes 9 of the first reaction tank A. have.
- a 9 mM urea aqueous solution was used as a sample solution, and a mixed solution containing 0.5 M hypobromite and 0.2 M sodium hydroxide was used as a reactant solution.
- the pump 5 and the pump 6 are operated, the sample solution and the reactant solution are introduced into the first reaction tank A from the ejection holes 9 of the first reaction tank A.
- a turbulent flow was generated in the first reaction tank A by the jet of the introduced solution, and the sample solution and the reactant solution were uniformly mixed to obtain a first mixed solution and nitrogen gas was generated.
- the obtained first mixed solution is introduced into the second reaction tank B from the ejection hole 11 through the ejection hole 10.
- a turbulent flow is generated in the second reaction tank B by the jet flow of the introduced first mixed solution, and the first mixed solution is further mixed to obtain a second mixed solution.
- the nitrogen gas introduced together with the first mixed solution from the ejection hole 11 of the second reaction tank B becomes fine bubbles by the jet flow of the first mixed solution and is dispersed in the second mixed solution. It was done.
- the obtained second mixed solution is further mixed in each reaction tank from the third reaction tank C to the seventh reaction tank G provided downstream of the discharge hole 12 of the second reaction tank B. Is done. At this time, nitrogen gas became finer bubbles with mixing in each reaction tank.
- FIG. 3 shows a chemiluminescence response waveform diagram obtained when both the sample solution and the reactant solution are introduced into the reaction vessel 2 while changing the sample solution and the reactant solution at the same flow rate.
- FIG. 4 shows a chemiluminescence response waveform diagram obtained when the concentration of the urea aqueous solution is changed.
- a chemiluminescence response waveform diagram used for calculation of the chemiluminescence lifetime ⁇ is shown in FIG. At this time, the chemiluminescence lifetime ⁇ was 0.9 seconds.
- FIG. 6 shows a correlation diagram between the urinary concentration and the chemiluminescence intensity.
- the inner radius of the ejection hole is less than 0.5 ⁇ 10 ⁇ 3 m.
- the time for which the solution having a flow rate of 0.667 ⁇ 10 ⁇ 6 m 3 / s stays in the reaction vessel 2 is about 0.22 seconds, which is necessary for the chemiluminescence reaction between urea and hypobromite ions. Since the time is about several hundreds of milliseconds, it is considered that almost all urea is consumed in the reaction vessel 2 and chemiluminescence occurs.
- Example 2 In Example 1, the reaction was carried out in the same manner as in Example 1 except that all the reaction tanks including the upper and lower surfaces of each reaction tank from the first reaction tank A to the seventh reaction tank G were made of acrylic resin.
- the chemiluminescence intensity generated by mixing the sample solution and the reactant solution in the container 2 was measured by the photodetector 24 to quantify the concentration of the sample solution.
- FIG. 7 shows a chemiluminescence response waveform diagram obtained when both the sample solution and the reactant solution are introduced into the reaction vessel 2 at the same flow rate. In FIG.
- Example 1 instead of using the reaction vessel 2 in which the respective reaction vessels from the first reaction vessel A to the seventh reaction vessel G were connected in series, a reaction vessel (diameter: 10 mm, height: 3 mm, A reaction vessel having only one (made of polyvinyl chloride) was used.
- a sample was prepared in the reaction vessel in the same manner as in Example 1 except that a mixed solution of 0.5 M sodium hypochlorite, 2 M sodium bromine, and 0.2 M sodium hydroxide was used as the reactant solution.
- the chemiluminescence intensity generated by mixing the solution and the reactant solution was measured by the photodetector 24, and the concentration of the sample solution was quantified.
- FIG. 8 shows a chemiluminescence response waveform diagram obtained when both the sample solution and the reactant solution were introduced into the reaction vessel at a flow rate of 20 ml / min.
- Example 3 In Example 1, instead of using the reaction vessel 2, each cylindrical reaction vessel from the first reaction vessel A to the seventh reaction vessel G (diameter: 3 mm, height 3 mm, made of polyvinyl chloride (black) ) was used in a straight line connected through a narrow passage (orifice, diameter 1 mm, length 3 mm).
- a transparent plate 26 (made of glass) having a thickness of 1 mm was disposed on the upper and lower surfaces of each of the reaction vessels A to G.
- a photo detector 24 photomultiplier tube
- a pinhole 27 equal to the diameter of each reaction vessel at a position facing the upper surface of the reaction vessel 25 is in contact with the upper surface. Arranged.
- FIG. 10 shows a chemiluminescence response waveform diagram obtained when the flow rate of each of the sample solution and the reactant solution is 20 ml / min (total flow rate is 40 ml / min). As can be seen from the chemiluminescence response waveform diagram of FIG.
- the chemiluminescence intensity showed the maximum value in the second reaction tank B, and thereafter decreased as the reaction tank became downstream. From this result, it can be seen that it is effective to connect two or more reaction vessels in series via a narrow passage (orifice) in order to obtain a strong chemiluminescence intensity.
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Abstract
Description
図1及び図2に示される試料溶液濃度測定装置1(縦:74mm、横:100mm、高さ:74mm)を用いて、試料溶液の濃度測定を行った。図1は試料溶液濃度測定装置1の一例を示した概略平面図であり、図2は前記装置1の概略側面図である。図1に示されるように、反応容器2(縦:40mm、横:40mm、高さ:5mm)は、第1の反応槽Aから第7の反応槽Gまでのそれぞれの円筒型反応槽(直径:3mm、高さ:3mm)が直列に接続された構成を有している。第1の反応槽Aから第7の反応槽Gまでの各反応槽が形成されたアクリル樹脂板の上面と下面にガラス板を配置した。これにより、前記各反応槽の上面と下面はガラスからなり、前記各反応槽の側面はアクリル樹脂からなる構成となった。第1の反応槽Aから第7の反応槽Gまでのそれぞれの反応槽には、噴出孔(内径:1mm)及び排出孔(内径:1mm)が設けられている。前記各反応槽の断面積(S1)と該噴出孔の断面積(S2)との断面積比(S1/S2)は9であった。第1の反応槽Aの排出孔10と第2の反応槽Bの噴出孔11とが細い通路を介して接続され、第2の反応槽Bの排出孔12の下流には更に第3の反応槽Cから第7の反応槽Gまでのそれぞれの反応槽が直列に接続されている。このとき、各反応槽における角度αは、反応槽Aは90度、反応槽Bは120度、反応槽Cは60度、反応槽Dは180度、反応槽Eは60度、反応槽Fは120度、反応槽Gは180度であった。図2に示されるように、反応容器2の上面と対向する位置に光検出器24である光センサモジュールを該上面と接するように取り付けた。
実施例1において、第1の反応槽Aから第7の反応槽Gまでの各反応槽の上面と下面を含め、全てアクリル樹脂製の反応槽とした以外は実施例1と同様にして、反応容器2内で試料溶液と反応剤溶液との混合により生じた化学発光強度を光検出器24により測定し、試料溶液の濃度を定量した。試料溶液と反応剤溶液の両方をそれぞれ同じ流量で反応容器2内に導入した際に得られた化学発光応答波形図を図7に示す。図7において、流量が10ml/minと小さい場合には、それ以上の流量(20ml/min、30ml/min、40ml/min)の場合に比べて光センサ出力の変動が激しく、化学発光の計測値が比較的不安定であった。このとき、反応槽内を目視により確認すると各反応槽内に残存する気泡が認められた。試料溶液と反応剤溶液との混合により発生した気泡の微細化が不十分であったと考えられる。したがって、前記発生した気泡によって化学発光反応が阻害されないようにするため、あるいは化学発光が散乱されないようにするためには、前記発生した気泡が微細化されるような墳流の速度、すなわち流量であることが望ましい。
実施例1において、第1の反応槽Aから第7の反応槽Gまでのそれぞれの反応槽が直列に接続された反応容器2を用いる代わりに、反応槽(直径:10mm、高さ:3mm、ポリ塩化ビニル製)を1つのみ有する反応容器を用いた。また、反応剤溶液として0.5Mの次亜塩素酸ナトリウム、2Mの臭素ナトリウム、及び0.2Mの水酸化ナトリウムの混合溶液を用いた以外は実施例1と同様にして、反応容器内で試料溶液と反応剤溶液との混合により生じた化学発光強度を光検出器24により測定し、試料溶液の濃度を定量した。試料溶液と反応剤溶液の両方をそれぞれ20ml/minの流量で反応容器内に導入した際に得られた化学発光応答波形図を図8に示す。
実施例1において、反応容器2を用いる代わりに、第1の反応槽Aから第7の反応槽Gまでのそれぞれの円筒型反応槽(直径:3mm、高さ3mm、ポリ塩化ビニル製(黒色))が細い通路(オリフィス、直径1mm、長さ3mm)を介して直線状に接続された反応容器25を用いた。各反応槽A~Gの上面と下面に厚さ1mmの透明板26(ガラス製)を配置した。図9に示されるように、反応容器25の上面と対向する位置に、各反応槽の直径と等しいピンホール27が設けられた光検出器24(光電子増倍管)を該上面と接するように配置した。実施例1と同様に、試料溶液として9mMの尿素を含む透析液を用い、反応剤溶液として0.5Mの次亜臭素酸と0.2Mの水酸化ナトリウムを含む混合溶液を用いた。リニアスライダー28により光検出器24を第1の反応槽Aから第7の反応槽Gまで移動させ、各反応槽A~Gにおける化学発光強度を測定した。試料溶液と反応剤溶液の各々の流量を20ml/min(合計流量40ml/min)とした場合に得られた化学発光応答波形図を図10に示す。図10の化学発光応答波形図から分かるように、化学発光強度は第2の反応槽Bにおいて最大値を示し、その後は下流の反応槽になるにつれて減少した。この結果から、強い化学発光強度を得るためには、細い通路(オリフィス)を介して反応槽が2つ以上直列に接続されていることが有効であることが分かる。
2 反応容器
3 試料溶液導入口
4 反応剤溶液導入口
5、6 ポンプ
7、8 配管
A 第1の反応槽
B 第2の反応槽
C 第3の反応槽
D 第4の反応槽
E 第5の反応槽
F 第6の反応槽
G 第7の反応槽
9 第1の反応槽の噴出孔
10 第1の反応槽の排出孔
11 第2の反応槽の噴出孔
12 第2の反応槽の排出孔
13 第3の反応槽の噴出孔
14 第3の反応槽の排出孔
15 第4の反応槽の噴出孔
16 第4の反応槽の排出孔
17 第5の反応槽の噴出孔
18 第5の反応槽の排出孔
19 第6の反応槽の噴出孔
20 第6の反応槽の排出孔
21 第7の反応槽の噴出孔
22 第7の反応槽の排出孔
23 排出口
24 光検出器
25 反応容器
26 透明板
27 ピンホール
28 リニアスライダー
Claims (8)
- 反応容器内で生じた化学発光強度を計測することにより試料溶液の濃度を定量する試料溶液濃度測定方法であって、
前記反応容器が、噴出孔及び排出孔を有する反応槽が2つ以上直列に接続されてなるものであり、
それぞれの前記反応槽の断面積(S1)と前記噴出孔の断面積(S2)との断面積比(S1/S2)が3以上であり、
試料溶液及び反応剤溶液を第1の反応槽の噴出孔から第1の反応槽に導入し、導入された溶液による噴流によって第1の反応槽内に乱流を発生させて該試料溶液と該反応剤溶液とを混合し、
得られた第1の混合溶液を第1の反応槽の排出孔を通じて第2の反応槽の噴出孔から第2の反応槽内に導入し、
前記第1の混合溶液による噴流によって第2の反応槽内に乱流を発生させて該第1の混合溶液を更に混合し、
前記反応容器が、第2の反応槽の下流に更に1つ又は複数の追加の反応槽が繰返し配置された構成を任意的に有していて、第2の反応槽内における上記混合と同様に混合溶液を更に混合してもよく、
前記反応容器内で生じた化学発光強度を計測することを特徴とする試料溶液濃度測定方法。 - 前記試料溶液が尿素を含む試料溶液である請求項1記載の試料溶液濃度測定方法。
- 前記試料溶液が透析廃液である請求項1又は2記載の試料溶液濃度測定方法。
- 前記反応剤溶液が次亜ハロゲン酸イオンを含む反応剤溶液である請求項1~3のいずれか記載の試料溶液濃度測定方法。
- 反応容器内で生じた化学発光強度を計測することにより試料溶液の濃度を定量する試料溶液濃度測定装置であって、
噴出孔及び排出孔を有する反応槽が2つ以上直列に接続されてなる反応容器を備え、
それぞれの前記反応槽の断面積(S1)と該噴出孔の断面積(S2)との断面積比(S1/S2)が3以上であり、かつその反応槽の少なくとも一部が透明部を有して該透明部の外側に光検出器が配置され、
試料溶液及び反応剤溶液を第1の反応槽の噴出孔から第1の反応槽に導入する手段を備え、導入された溶液による噴流によってそれぞれの前記反応槽内に乱流を発生させて、該試料溶液と該反応剤溶液とを混合することにより前記反応容器内で生じた化学発光強度を計測することを特徴とする試料溶液濃度測定装置。 - 前記噴出孔の軸と前記排出孔の軸との交わる角度αが15度以上165度以下となる反応槽を少なくとも1つ有する請求項5記載の試料溶液濃度測定装置。
- 尿素濃度を定量する請求項5又は6記載の試料溶液濃度測定装置。
- 請求項7記載の試料溶液濃度測定装置により透析廃液中の尿素濃度を定量する人工透析装置。
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EP3376212A1 (en) | 2017-03-17 | 2018-09-19 | Ricoh Company Ltd. | Chemiluminescence analyzer, blood purification apparatus, and blood purification system |
JP2018155733A (ja) * | 2017-03-17 | 2018-10-04 | 株式会社リコー | 化学発光分析装置、血液浄化装置、及び血液浄化システム |
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