JP2011007666A - Sample liquid dilution method and device of the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a sample liquid dilution device that is inexpensive, is easily operated, and automatically performs accurate sample liquid dilution.SOLUTION: A sample liquid and diluent are mixed by using a device communicating a pair of syringes 5 and 6 with each other via a communication passage 25 between syringe cylindrical bodies, injecting the sample liquid and a diluent into a diluted/stirred liquid storage part constituted by both the syringes 5 and 6 and the communication passage 25 between the syringe cylindrical bodies, and making the communication passage 25 between the syringe cylindrical bodies an orifice, and through which, reciprocating and moving both liquids into both syringes 5 and 6.

Description

  The present invention relates to a sample liquid diluting apparatus for diluting a sample liquid to a state necessary for a desired analysis process.

  Conventionally, when using an analytical instrument that automatically performs high-performance liquid chromatography (HPLC) or surface plasmon resonance (SPR method), sample solution adjustment involving dilution operation is required.

  As a sample liquid dilution method by manual operation, a method using a micropipette as a measuring liquid collecting means and a vortex mixer as a mixing means is generally used, but a whole pipette and a mixing means are used as the measuring liquid collecting means. In addition, a simple sample solution dilution method using a volumetric flask can perform more quantitative dilution with the highest accuracy.

  As a sample liquid dilution method in a laboratory automation system (hereinafter LAS), pipetting by a dispenser is performed, and mixing by a vortex mixer, a shaker or the like is used (for example, Patent Documents 1 and 2).

JP-A-6-142482 JP 2000-146784 A

  However, manual operation in the conventional technology as described above requires personal skills, and cannot be said to be highly accurate by anyone, and there is a problem that human error is likely to occur.

  In addition, the dispenser mounted in LAS generally has a liquid collection volume of about 10 μl to 1000 μl, but the repeated dispensing accuracy is about 0.5% or less, but the accuracy exceeding this volume range is high. Dispensing is difficult, and the highly accurate dilution rate per one time by LAS is about 100 times. Usually, enzyme immunoassay (ELISA method) and surface plasmon resonance method (SPR method) that require dilution are tens of thousands. There is a problem that dilution more than double is essential, and in order to achieve this dilution ratio in the LAS, multiple dilutions are required, resulting in a reduction in dilution accuracy.

  Furthermore, although many fully automatic ELISA systems using LAS are currently sold, they are very expensive and high accuracy cannot be expected for the reasons described above.

  An object of the present invention is to provide a sample liquid dilution method and an apparatus thereof that can automatically perform high-precision sample liquid dilution that is inexpensive, easy to operate, and in view of the above-described problems.

  The invention according to claim 1 for solving the conventional problems as described above and achieving an intended purpose uses an apparatus in which a pair of syringes are communicated with each other via a communication path between syringe barrels. The sample liquid and the diluent are injected into a dilute stirring liquid storage portion configured by the syringe and the syringe cylinder communication path, and the syringe cylinder communication path is used as an orifice, and the both liquids are passed through the syringes. The sample liquid dilution method is characterized in that the sample liquid and the diluent are mixed by reciprocating.

  The invention according to claim 2 is a cylindrical first syringe cylinder whose one end is closed, and a plunger which is reciprocally inserted into the syringe cylinder and is forced to reciprocate manually or by a reciprocating drive mechanism. A first syringe comprising: a cylindrical second syringe cylinder having at least one end closed; a second syringe comprising a plunger or a piston reciprocally inserted into the syringe cylinder; and the two syringes A communication path between syringe cylinders for communicating between the closed ends of the syringe cylinders, a sample injector for quantitatively supplying a sample into one of the two syringes, and a diluent to be supplied into one of the two syringes A diluent supply means, and the sample solution is supplied from the sample injector into the liquid storage portion constituted by the syringe and the communication path between the syringe barrels. The sample is characterized in that the diluent is injected from each means, and the plunger is reciprocated to reciprocate the two solutions into the syringes through the communication path between the syringe barrels. In the liquid diluter.

  According to a third aspect of the present invention, in addition to the configuration of the second aspect, a supply / drain liquid path is connected to the communication path between the syringe cylinders, and the dilution liquid supply is connected to the supply / drain liquid path via a switching valve. The dilute supply path from the means and the dilute mixture discharge path can be switched and communicated.

  According to a fourth aspect of the present invention, in addition to the configuration of any one of the second and third aspects, the second syringe is configured such that the syringe head side of the second syringe cylinder partitioned by the piston is disposed on the liquid storage portion. The piston is always urged toward the syringe head by the return pressurizing means on the back side.

  According to a fifth aspect of the present invention, in addition to the configuration of the fourth aspect, the return pressurizing means includes a pneumatic pressurizing device in which the back side of the piston is a sealed space and the sealed space is filled with pressurized air. It is to be.

    In the present invention, a device in which a pair of syringes are communicated with each other via a communication path between syringe barrels, and the sample liquid and The diluent is injected, and the sample liquid and the diluent are mixed by reciprocating the two solutions into the syringe through the syringe cylinder communication path as an orifice. Even when the mixed liquid with the sample liquid flows into one of the syringes from the orifice, a violent vortex occurs, and even if the sample liquid volume is very small and the dilution ratio is high, uniform mixing is possible. The state can be easily obtained in a short time, and the dilution accuracy is improved.

  The present invention also includes a first cylindrical syringe barrel whose one end is closed and a plunger which is reciprocally inserted into the syringe barrel and is forced to reciprocate manually or by a reciprocating drive mechanism. 1 syringe, a cylindrical second syringe cylinder closed at least at one end, a second syringe consisting of a plunger or piston reciprocally inserted into the syringe cylinder, and syringe cylinders of both syringes A communication path between syringe cylinders for communicating between the closed ends, a sample injector for quantitatively supplying a sample into one of the two syringes, and a diluent supply for supplying a diluent into one of the two syringes Means for supplying the sample liquid from the sample injector into the liquid storage part constituted by the syringe and the communication path between the syringe barrels, from the diluent supply means. By injecting each of the diluents and reciprocating the plunger, the two liquids are reciprocated into the syringes through the communication path between the syringe barrels to be mixed. High precision dilution is achieved with low equipment.

  Further, in the present invention, a supply / discharge liquid path is communicated with the syringe cylinder communication path, and the dilution liquid supply path and the diluted mixed liquid discharge from the dilution liquid supply means are connected to the supply / discharge liquid path via a switching valve. By making it possible to switch and communicate with the path, the flow path configuration is simplified, and the manufacture of the device is facilitated.

  Furthermore, in the present invention, the second syringe uses the syringe head side of the second syringe cylinder partitioned by the piston as the liquid storage portion, and the piston is always on the syringe head side by the return pressurizing means on the back side. By energizing the plunger, the liquid mixture is efficiently moved between the syringes by an operation of drinking the plunger of one syringe.

  Further, according to the present invention, the structure is simplified by using a pneumatic pressurizing device in which the back side of the piston is a sealed space and the sealed space is filled with pressurized air as the return pressurizing means. Can with equipment.

It is a channel figure showing the outline of the sample dilution device concerning the present invention. It is a partial flow path figure which shows the sample liquid measurement state of a sample injector same as the above. It is a flow chart which shows a sample solution injection state same as the above. It is a flow-path figure which shows a dilution liquid suction state same as the above. It is a flow path figure which shows the diluted liquid discharge state same as the above.

  Next, embodiments of the present invention will be described based on examples shown in the drawings. In the figure, 1 is a mixing and stirring unit, 2 is a sample injector, and 3 is a diluent supply unit.

  The mixing and stirring unit 1 has a pair of syringes including a first syringe 5 and a second syringe 6. The first syringe 5 includes a cylindrical first syringe cylinder 10 and a plunger 11. The plunger 11 includes a piston 11a that partitions the first syringe cylinder 10 while maintaining airtightness, and an operation shaft portion 11b having a distal end fixed thereto.

  The second syringe 6 includes a cylindrical second syringe cylinder 13, a piston 14 that partitions the interior thereof while maintaining airtightness, and a return actuator for biasing the piston 14 toward the syringe head 20 described later. It is comprised from the pneumatic chamber 15 which is a pressure means. The pneumatic chamber 15 is filled with compressed air, and the back surface of the piston 14 is constantly pressurized.

  Both syringes 5 and 6 are integrated with a common syringe head 20 at the front end of the first syringe cylinder 10 and the second syringe cylinder 13 and integrated with a common flange block 21 at the rear end. Yes. The rear end of the first syringe cylinder 10 is opened to the back surface of the flange block 21, and the plunger 11 is inserted through the opening. The rear end of the second syringe cylinder 13 is a sealed space closed by a plug 22. The plug 22 is provided with an air inlet 23 with a check valve.

  In this embodiment, air pressure is used as the return pressurizing means for pressing the back surface of the piston 14, but an elastic material such as a coil spring may be used.

  The syringe head 20 is formed with a syringe-cylinder communication passage 25 that allows communication between the closed ends of the syringe cylinders 10 and 13. The communication path 25 between the syringe cylinders is formed to have a smaller diameter than the inner diameters of the syringe cylinders 10 and 13 and constitutes an orifice (throttle channel) that allows the syringe cylinders 10 and 13 to communicate with each other. The syringe cylinder communication passage 25 and the syringes 5 and 6 serve as a diluted stirring liquid storage section.

  A central portion of the syringe cylinder communication passage 25 can be opened and closed by an opening / closing valve 26, and a supply / drainage channel 27 is communicated with the first syringe cylinder 10 side. A sample injection path 28 communicates with the side. The sample injection path 28 may communicate directly with the closed end portion of the second syringe cylinder 13 instead of the syringe cylinder communication path 25.

  The sample injector 2 is rotatably accommodated in a stator 30 and has a rotor 31. The rotor 31 is provided with a measuring loop 32, and both ends thereof are opened to the outer periphery of the rotor. The stator 30 is provided with two sample measurement ports 32a and 32b and sample delivery ports 33a and 33b, which are matched at both ends of the measurement loop 32, for injecting a sample into one of the sample measurement ports 32a. The sample syringe 34 is communicated, and the surplus sample discharge channel 36 communicating with the surplus sample storage container 35 is communicated with the other port 32b.

  A sample delivery syringe 37 communicates with one port 33a for sample delivery, and a sample injection path 28 communicates with the other port 33b. A drainage path 40 communicating with the drainage container 39 is communicated with the sample injection path 28 via a three-way switching valve 38.

  In the state shown in FIG. 2, the sample injector 2 configured as described above injects the sample liquid by the sample syringe 34 to fill the sample liquid in the measuring loop 32, and then rotates the rotor to the state shown in FIG. By injecting the diluent from the syringe 37, the sample solution in the measuring loop 32 is injected into the second syringe cylinder 13 through the sample injection path 28.

  The three-way switching valve 38 is used when the inside of the measurement loop 32 is washed by injecting the sample liquid into the second syringe cylinder 13 and then flowing the diluent into the waste liquid container 39 in the delivery syringe 37. To do.

  The diluent supply unit 3 is provided with a diluent container 41, and has a diluent supply path 42 that leads the diluent in the container to the supply / discharge liquid path 27 of the mixing and stirring unit 1 described above through the degasser 46. . The diluent supply path 42 is in communication with one switching port of the three-way switching valve 43 so as to be switched to and communicated with the supply / discharge liquid path 27. The other switching port of the three-way switching valve 34 communicates with a diluted mixed solution discharge path 45 that leads to a diluted liquid container 44 for storing diluted mixed liquid.

  A sample solution dilution method using the apparatus configured as described above will be described. First, as shown in FIG. 3, the plunger 11 of the first syringe 5 and the piston 14 of the second syringe 6 are pushed into the syringe head side most. At this time, the three-way switching valve 38 is operated to the side where the sample injection channel 28 and the port 33b communicate with each other, and the on-off valve 26 is closed.

  In this state, the delivery syringe 37 is operated to inject the sample liquid previously stored in the measuring loop 32 into the second syringe 6. After the injection is completed, the switching valve 38 is operated to close the sample injection path 28.

  Next, as shown in FIG. 4, the three-way switching valve 43 is operated in a direction in which the plunger 11 of the first syringe 5 is retracted by directing the diluent channel 42 side to the side where the diluent channel 42 communicates with the supply / drain channel 27. The diluted solution in the liquid container 41 is sucked into the first syringe 5.

  In this way, after the sample liquid and the diluted liquid are accommodated in the first and second syringes 5 and 6 so as to have a desired dilution ratio, as shown in FIG. The on-off valve 26 is opened while operating to the side where the passage 27 is closed.

  In this state, the diluent 11 in the first syringe 5 is injected into the second syringe 6 by pushing the plunger 11. At this time, the communication path 25 between the syringe cylinders becomes an orifice, and the symptom is generated because the syringe cylinder enters the second syringe cylinder and rapidly expands, and is mixed with the sample liquid in the second syringe 6 and stirred. Become. At this time, the piston 14 moves backward against the air pressure on the back surface thereof.

  After the diluted solution is moved into the second syringe 6 in this way, the mixed solution in the second syringe 6 is moved into the first syringe 5 by operating the plunger 11 in the backward direction. Stirring is performed when flowing into the first syringe cylinder 10 from the communication path 25 formed. By performing the operation of the plunger 11 a plurality of times, the diluted solution and the sample solution are uniformly mixed.

  After this mixing operation is completed, the plunger 11 is moved backward to move the mixed liquid to the inside of the first syringe 5, and the open / close valve 26 is closed as shown in FIG. Is operated so that the supply / discharge liquid path 27 leads to the diluted mixed liquid discharge path 44, and then the plunger 11 is moved in the pushing direction. As a result, the liquid mixture in the first syringe 5 is sent out into the diluted liquid container 43.

  The pushing of the diluted liquid mixture into the diluted liquid container 43 is performed by operating the three-way switching valve 43 in a state where the mixed liquid is moved into the second syringe 6, so You may make it extrude by.

  In this embodiment, the plunger 11 is manually operated. For example, the plunger may be operated by using an electric drive mechanism in which a stepping motor and a crank mechanism are combined. .

DESCRIPTION OF SYMBOLS 1 Mixing stirring part 2 Sample injector 3 Diluent supply part 5 1st syringe 6 2nd syringe 10 1st syringe cylinder 11 Plunger 11a Piston 11b Operation shaft part 13 2nd syringe cylinder 14 Piston 15 Pneumatic chamber 20 Syringe head 21 Flange block 22 Plug body 23 Air inlet 25 Communication path 26 between syringe cylinders On-off valve 27 Supply / drain liquid path 28 Sample injection path 30 Stator 31 Rotor 32 Metering loops 32, 32b Sample metering ports 33a, 33b Sample delivery Port 34 Sample syringe 35 Surplus sample storage container 36 Surplus sample discharge flow path 37 Sample delivery syringe 38 Three-way switching valve 40 Drainage path 41 Diluent container 42 Diluent supply path 43 Three-way switching valve 44 Diluted liquid container 45 Dilution mixing Liquid discharge passage 46 Degasser

Claims (5)

  Using a device in which a pair of syringes are communicated with each other via a communication path between syringe cylinders, the sample liquid and the dilution liquid are injected into the diluted stirring liquid storage portion constituted by the syringes and the syringe cylinder communication path. The sample liquid dilution method is characterized in that the sample liquid and the diluting liquid are mixed by reciprocating the both liquids into the syringes through the syringe cylinder communication path as an orifice.   A first syringe consisting of a cylindrical first syringe cylinder closed at one end, and a plunger that is reciprocally inserted into the syringe cylinder and is forced to reciprocate manually or by a reciprocating drive mechanism; and at least one end A cylindrical second syringe cylinder closed, a second syringe composed of a plunger or a piston reciprocally inserted into the syringe cylinder, and the syringe cylinder closed ends of the two syringes communicating with each other A syringe cylinder communication path, a sample injector for quantitatively supplying a sample into one of the two syringes, and a diluent supply means for supplying a diluent into one of the two syringes, Sample liquid from the sample injector and dilution liquid from the dilution liquid supply means in the liquid storage part constituted by the communication path between both syringes and the syringe barrel, respectively. A sample liquid diluting apparatus, wherein the two liquids are reciprocated into the syringes through the communication path between the syringe cylinders and mixed by injecting and reciprocating the plunger.   A supply / discharge liquid path is connected to the communication path between the syringe barrels, and the dilution liquid supply path from the dilution liquid supply means and the diluted mixed liquid discharge path can be switched and communicated to the supply / discharge liquid path via a switching valve. The sample solution diluting device according to claim 2.   The second syringe uses the syringe head side of the second syringe cylinder partitioned by the piston as a liquid storage portion, and the piston is always urged toward the syringe head side by a return pressurizing means. The sample solution diluting device according to claim 2 or 3. 5. The sample liquid diluting device according to claim 4, wherein the returning pressurizing means is a pneumatic pressurizing device in which the back side of the piston is a sealed space and the sealed space is filled with pressurized air.

Images