JPH063440U - HPLC sample filling device - Google Patents

Abstract

(57) [Summary] [Objective] The entire amount of the sample in the sample container is packed into the HPLC column through a 6-way switching valve with a loop without air inclusion. [Structure] A sample container, an HPLC pump, a column, and a sample transfer pump for performing an extrusion / suction operation are connected to each port of a looped 6-way switch valve through a passage, and the sample transfer pump is a 6-way switch valve. The liquid in the loop is pushed until it reaches the sample container, the air between the 6-way switching valve and the sample container is pushed out to the sample container, and then the sample is sucked from the sample container into the loop. In addition, a liquid detection sensor is installed in the passage between the 6-way switching valve and the sample container,
It is detected that the entire amount of the sample in the sample container is filled in the loop of the 6-way switching valve.

Description

[Detailed description of the device]

[0001]

[Industrial applications]

 The present invention relates to a device for filling a sample into an HPLC column used for an automatic compound synthesizer, etc., and particularly, to enable the column to be automatically filled with the entire amount of the sample in the sample container while preventing air from being mixed. It is a thing.

[0002]

[Prior art]

 Conventionally, in this type of HPLC sample filling device, as shown in FIG. 3, a sample container 1 is connected to a 6-way switching valve 2 equipped with a loop 2a via a pipe 3, and the 6-way switching valve 2 is connected to the 6-way switching valve 2. A sample metering suction pump 5 and a column 6 from the HPLC pump 4 are connected via pipes 7, 8 and 9, respectively.

The operation of filling the sample in the sample container 1 into the column 6 is as follows. First, with the 6-way switching valve 2 at the position shown in the figure, the developing solvent from the HPLC pump 4 is passed through the loop 2 a of the 6-way switching valve 2 to the column. Supply to 6. Next, the 6-way switching valve 2 is switched to open the passages for the sample fixed-quantity suction pump 5, the loop 2a, and the sample container 1, and the sample container 1 is filled with a predetermined volume of sample in the loop 2a. Next, the 6-direction switching valve 2 is switched, and the HPLC pump 4 supplies the sample filled in the loop 2a to the column 6.

[0004]

[Problems to be solved by the device]

 In the above-mentioned conventional apparatus, the air in the pipe 3 between the sample container 1 and the loop 2a is introduced into the loop 2a and filled in the column 6 together with the sample. When air is mixed in the column 6, the column 6 is deteriorated, the chromatographic detection peak is disturbed, and it becomes impossible to adopt the automatic peak division method by the computer.

In addition, since a predetermined fixed amount of sample is sampled from the sample container by the suction pump and packed in the column, it is good for analysis, but the total amount of the sample in the sample container is packed in the column 6. When performing separation and purification, loss of sample is unavoidable. Therefore, when the column is filled with the entire amount of the sample in the sample container, a method of filling the column with the fixed amount of the sample is repeated to divide the sample into the column.

In this case, it is necessary to accurately know the total amount of the sample in advance, and the number of times of the dividing operation needs to be calculated in advance, which is a troublesome task. Also, in the case of divided filling, unless the sample has a volume that is a multiple of the set pump volume, it is not possible to fill only the sample into the loop without air inclusion.

On the other hand, when the volume of the quantitative pump is increased so that the total amount of the sample is introduced into the loop at one time, it is difficult to fill the total amount of the sample into the column depending on the accuracy of the quantitative pump. It was

The present invention has been made in view of the above problems, and an object of the present invention is to enable the column to be filled with the entire amount of the sample in the sample container without the inclusion of air.

[0009]

[Means for Solving the Problems]

 In order to achieve the above-mentioned object, the present invention connects a sample container, an HPLC pump, a column and a sample transfer pump for performing extrusion / suction operation to each port of a 6-way switching valve with a loop through a passage, Use the sample transfer pump to push out the liquid in the loop of the 6-way switching valve until it reaches the sample container, push the air between the 6-way switching valve and the sample container into the sample container, and then put the sample in the loop from the sample container. An HPLC sample filling device having a suction structure is provided.

A liquid detection sensor is installed in the passage between the 6-way switching valve and the sample container so that it is possible to detect that the entire amount of the sample in the sample container is filled in the loop of the 6-way switching valve. There is.

Further, the capacity of the passage between the 6-way switching valve and the sample transfer pump is set to be equal to or larger than the loop capacity.

As the sample transfer pump, it is preferable to use a syringe pump that pushes and pulls a piston into a fixed tube that communicates with the tube. Further, it is preferable that a stepping motor is used as a drive source for operating the piston of the syringe pump, and the liquid is slowly transferred by the sample transfer pump.

[0013]

[Action]

 According to the sample filling device having the above-described configuration, the developing solvent filled in the loop of the 6-way switching valve is pushed out by the sample transfer pump until it reaches the sample container. Therefore, the air between the loop and the sample container is evacuated. Are extruded to the sample container side. After that, when the sample in the sample container is sucked into the loop, the sample can be packed into the loop and from the loop to the column without air inclusion.

Further, since the liquid detection sensor is installed in the passage between the sample container and the 6-way switching valve, the liquid does not pass through the passage, so that the total amount of the sample in the sample container is the loop of the 6-way switching valve. Can be detected. Therefore, the total amount of the sample can be packed into the column from the loop in one operation without knowing the total amount of the sample in the sample container in advance.

[0015]

【Example】

 Hereinafter, the present invention will be described in detail with reference to the embodiments shown in the drawings. As shown in FIG. 1, a tube 11 is connected to a supply port 10a formed on the bottom surface of the sample container 10, and the tube 11 is connected to a port of a 6-way switching valve 12. A liquid detection sensor 13 that detects whether or not a sample is flowing in the tube 11 is installed in the middle of the tube 11.

A tube 14 is connected to a port of the 6-way switching valve 12, the tube 14 is connected to tubes 16 and 17 via a 3-way solenoid valve 15, and the tube 16 is connected to a syringe pump 18. At the same time, the tube 17 is vacuum pumped and connected to the drain tank 19.

Further, the ports of the 6-way switching valve 12 are communicated with each other via the loop 12 a, the HPLC pump 21 is connected to the port via the tube 20, and the column 23 is connected to the port via the tube 22. is doing. A detector 25 is connected to the column 23 via a tube 24. Each of the tubes 11, 14, 16, 17, 20, 22, 24 constituting the above passage is a Teflon tube.

In the illustrated state, the 6-way switching valve 12 has a port, a port, and a port that are connected by a solid line, and communicates with each other. When the switching operation causes the port to rotate in the clockwise direction, Portions indicated by dotted lines are in communication, that is, ports, ports, and ports are in communication with each other.

The syringe pump 18 has a structure shown in FIG. 2, in which one end of the piston 27 is slidably fitted in one end opening of the fixed tube 26 and the tube 16 is connected to the other end of the fixed tube 26. In accordance with the movement of the piston 27, the tube 16 connected to the fixed tube 26 pushes the liquid to the 6-way switching valve 12 or sucks the liquid into the tube 16.

The fixed tube 26 has a first fixed plate 30 fixed to the outer peripheral surface thereof connected to a second fixed plate 31, and the second fixed plate 31 is erected on a fixed base 32 so that the fixed tube 26 is fixed. It is fixed in position. The piston 27 is movably penetrated through a guide hole 31a formed in the second fixing plate 31. Further, the first moving plate 34 is fitted into the tip neck portion 27 a of the piston 27, and the first moving plate 34 is connected to the second moving plate 35. The second moving plate 35 is erected on the moving base 36, and the second and first moving plates 34, 35 move in accordance with the movement of the moving base 36 to push the piston 27 into the fixed pipe 26. To or to the pulling direction.

The moving base 36 and the fixed base 32 are attached to a screw rod 40 axially supported between bases 38 and 39 arranged opposite to each other and guide rails 41 on both sides thereof, and the fixed base 32 is fixed by a screw means 32c. It is fixed to the guide rail 41, and the screw rod 40 is loosely inserted in the hole 32a formed in the fixing base 32. On the other hand, the movable table 36 is slidably fitted to the guide rail 41, and the threaded holes 36a formed therein are screwed into the threaded rods 40 so as to reciprocate according to the rotation of the threaded rods 40. . One end of the screw rod 40 is projected from the base 38 and is rotated by a stepping motor 43 fixed to a motor mounting plate 42.

In the syringe pump 18, the liquid can be slowly transferred by pushing or pulling the piston 27 by driving the stepping motor 43.

The volume of the loop 12 a of the above-mentioned 6-way switching valve 12 is A, the volume of the tube 14 from the 6-way switching valve 12 to the switching valve 15 is B, the volume of the syringe pump 18 is C, and the sample to be put in the sample container 10 When the total amount is D, D ≦ A ≦ B ≦ C is set. In this embodiment, A, B and D are all set to 30 ml. The cylinder pump has a capacity of 50 ml and the tube 11 has a capacity of 5 ml.

The liquid detection sensor 13 is connected to a computer (not shown), and the computer controls the driving of the 6-way switching valve 12, the stepping motor 43 for driving the syringe pump 18, and the HPLC pump 21. There is.

The sample container 10 is connected to the cleaning liquid reservoir 50 via a tube 51 and a switching valve 52.

Next, the operation of the above device will be described. First, the sample (reagent) to be purified is put in the sample container 10. The sample is 30 ml. Then, the HPLC pump 21 is driven to supply the developing solvent from the HPLC pump 21 to the six-way switching valve 12 through the tube 20, and the loop 12a is filled with the developing solvent from the port through the port. And the detector 25 is operated.

After that, the 6-way switching valve 12 is switched, and the dotted line position in FIG. 1 is set as a communication path, and the communication in the solid line position is blocked. Therefore, the port → port → loop 12a → port → port are communicated, and the line of syringe pump 18 → tube 16 → tube 14 → loop 12a → tube 11 → sample container 10 is opened.

Next, the stepping motor 43 of the syringe pump 18 is driven, and the piston 27 is pushed into the fixed tube 26 side to operate. By this operation, the air in the tubes 14 and 16 is pushed toward the loop 12a side, and the developing solvent filled in the loop 12a passes through the tube 11 and is pushed out until it reaches the sample container 10. By the above operation, the air in the tube 11 between the 6-way switching valve 12 and the sample container 10 is pushed out to the sample container 10 side.

Next, the stepping motor 43 of the syringe pump 18 is rotated in the reverse direction to pull the piston 27 to operate. By the above operation, the developing solvent in the tube 11 and the loop 12a is sucked to the tubes 14 and 16 side, and the sample in the sample container 10 is slowly transferred to the loop 12a.

The liquid detection sensor 13 detects whether or not the sample in the sample container 10 has passed through the tube 11, and when it is detected that no liquid has passed, the entire amount of the sample in the sample container 10 becomes a loop 12a. It can be confirmed that it was transferred. When the computer determines that the entire amount of the sample is supplied to the loop 12a based on the detection signal from the liquid detection sensor 13, the 6-way switching valve 12 is switched to return to the state shown in FIG.

By the above switching operation, the lines of the HPLC pump 21, the tube 20, the loop 12a, the tube 22, the column 23, the tube 24, and the detector 25 are opened, and the total amount of the sample filled in the loop 12a is stored in the column 23. Is filled.

As described above, since the entire amount of the sample is packed and discharged without mixing air in the column 23, the purification is performed by the peak division method in which the fraction is switched for each peak according to the detection value from the detector 25. You can

After completion of the chromatography, the stepping motor 43 of the syringe pump 18 is driven to push the piston 27 to return it to the initial setting position. The developing solvent is returned to the sample container, discarded during washing, and prepared for the next purification.

In the above embodiment, a syringe pump is used to push out the developing solvent filled in the loop to the side of the sample container 10 and push out the air in the tube 11, but the developing pump in the loop is used. The pump is not limited to the syringe pump as long as the solvent is pushed out to the sample container side through the tube 11 by a certain amount, and other suitable extrusion pump can be used. However, the use of a syringe pump is preferable because a certain amount of liquid can be slowly transferred.

[0035]

[Effect of device]

 As is clear from the above description, according to the HPLC sample filling apparatus of the present invention, the developing solvent filled in the loop of the 6-way switching valve is pushed out to the tip of the sample container by the pump, and thus the 6-way switching valve is used. The air in the tube between the sample containers can be pushed out to the sample container, and subsequently, when the sample in the sample container is supplied to the loop, it can be filled without air inclusion, and the column is filled from the loop. It is possible to reliably prevent air from being mixed into the sample.

Therefore, since there is no disorder in the chromatographic detection peak, the fractions can be switched and purified by an automatic division method by a computer.

Further, since the liquid detection sensor is arranged in the tube for transferring the sample from the sample container to the loop, the entire amount of the sample in the sample container can be surely filled in the loop. Therefore, it is possible to eliminate the loss of the sample when performing the fractional purification.

[Brief description of drawings]

FIG. 1 is a schematic view showing the entire device of the present invention.

FIG. 2 is an exploded perspective view of a syringe pump used in the above device.

FIG. 3 is a schematic diagram of a conventional device.

[Explanation of symbols]

 10 sample container 12 6-direction switching valve 12a loop 13 liquid detection sensor 18 syringe pump 21 HPLC pump 23 column 25 detector 11, 14, 16, 17, 20, 22, 24 tube

Claims (5)

[Scope of utility model registration request] 1. A sample container, an HPLC pump, a column, and a sample transfer pump for performing an extrusion / suction operation are connected to each port of a 6-way switching valve with a loop through a passage,
With the sample transfer pump, the liquid in the loop of the 6-way switching valve is pushed out until it reaches the sample container, and the air between the 6-way switching valve and the sample container is pushed out to the sample container, and then the sample is moved from the sample container into the loop. HP configured to suck
LC sample filling device. 2. A liquid detection sensor is installed in the passage between the 6-way switching valve and the sample container so that it is possible to detect that the entire amount of the sample in the sample container has been filled in the loop of the 6-way switching valve. The device of claim 1, wherein 3. The apparatus according to claim 1, wherein the capacity of the passage between the 6-way switching valve and the sample transfer pump is set to be equal to or larger than the loop capacity. 4. The apparatus according to claim 1, wherein a syringe pump that pushes and pulls a piston into and out of a fixed tube that is in communication with a tube is used as the sample transfer pump. 5. The apparatus according to claim 4, wherein a stepping motor is used as a drive source for operating the piston of the syringe pump.