JP2001059839A - Automatic refining device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To prevent the loss of a sample by pumping liquid from the sample vessel of a sample to a column by one fixed quantity injector and from the storage vessel of a developing solvent to the column and sharing a pumping passage. SOLUTION: The fixed quantity injector 20 of a sample filling part provides a flow inlet 20a and a flow outlet 20b on the head member 42 of the upper end of a syringe 30 housing a piston 34, and a groove 20c connecting them is opened on the lower face of the head member 42. When the piston 34 is lifted to be contacted on the lower face of the head member 42, the lower openings of the flow inlet 20a, the flow outlet 20b and the groove 20c are closed, and the flow inlet 20a and the flow outlet 20b form a short-circuited flow passage 20d short-circuiting through the groove 20c. The piston 34 is lowered to suck the sample in the inside of the syringe 30, a fixed quantity of the sample is stored, the piston 34 is lifted to push out the sample, an air layer is removed by the changeover of a liquid surface sensor and a rotary valve, after only the sample is injected into a column, the short-circuited passage 20d is passed at the position of the upper limit of the piston 34, and a developing solvent is injected into the column.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an automatic purification apparatus for purifying a sample in an organic compound synthesis step,
It can be used as an automatic synthesis device in combination with a device for performing a production reaction step, or can be used as a single device.

[0002]

2. Description of the Related Art The process of synthesizing an organic compound can be broadly divided into a production reaction process and a purification process. After a compound is synthesized in the production reaction process, a purification process is always required, and sometimes the purification process is performed more than the production reaction process. In some cases, the step is more important, and in many cases, the time required for the purification step is longer than the time for the production reaction step.

[0003] The types of purification methods include extraction, recrystallization, distillation, sublimation, chromatography and the like. Among them, the purification method by chromatography using a column is most widely used due to its separation ability, high efficiency and the like. ing.

[0004]

However, in the above-described purification method by chromatography, the experience of the experimenter is an important factor, the operation is complicated, the time required is long, and the time constraint of the experimenter is long. There was a problem.

[0005] In response to the above problems, the present applicant has previously incorporated a purification apparatus using a chromatography method into an automatic synthesis apparatus, and integrated a synthesis reaction step and a purification step into a series of synthesis steps to synthesize organic compounds by computer control. We provide equipment that can be fully automated.

[0006] The conventional purification apparatus has the advantage that it can be fully automated as described above. However, since a loop method is used as a path for sending the sample to the column, the sample is diffused when a large amount of sample is injected. However, there is a problem that the purification capacity is reduced. Further, since the sample is extruded into the column by the developing solvent, the sample is diluted with the developing solvent in a long path, and there is a problem that the sample separating ability is deteriorated. Further, since different liquid paths are usually provided separately, there is a problem that the structure is complicated and the number of parts is increased.
Furthermore, when bubbles are mixed in the effluent from the column, since no bubble removing means is provided, if liquid is sent to the detector while containing bubbles, it causes noise and a true peak cannot be obtained. There was a problem.

[0007] The present invention has been made in view of the above problems, and an object of the present invention is to provide a purification apparatus using a column chromatography method, in which the above various problems are not solved.
In particular, it is an object of the present invention to provide an automatic purification apparatus that can be used alone and can be used as a fully automatic synthesis apparatus in combination with a production reaction apparatus.

[0008]

To achieve the above object, according to the present invention, a driving unit and a control unit are provided, wherein the driving unit includes a developing solution mixing / sending unit and a sample supplying unit. , A sample filling section, a column section, a bubble removing section, an effluent component detecting section, and an effluent dividing section. The developing liquid mixing and sending section and the sample supplying section are each provided via a flow line provided with an electromagnetic valve. Connected to a first rotary valve, and the first rotary valve is connected to a quantitative injector provided in the sample filling section via a flow line.
The quantitative injection device is sequentially connected to the second and third rotary valves via a flow line, the third rotary valve is connected to the column portion, and the developing solution is sent from the quantitative injection device to the column portion after the sample is sent to the column portion. To the column portion, and the effluent from the column portion is sent to the effluent dividing portion through a flow line provided with the bubble bleed portion and the component detecting portion of the effluent, and In addition, the present invention provides an automatic refining apparatus characterized in that the operation of the solenoid valve and the rotary valve is controlled by a signal from the control unit.

The metering injector provided in the sample filling section is provided with an inlet and an outlet having upper and lower openings on both sides of a head member attached to an upper end of a syringe accommodating a piston so that the piston can move up and down, and connects the inlet and the outlet. When the piston comes into contact with the lower surface of the head member at the ascending limit, the inflow port, the outflow port and the lower surface opening of the groove are closed, and the inflow port and the outflow port are closed. Forms a continuous short-circuit channel through the groove, aspirates the sample inside the syringe by lowering the piston, temporarily collects a certain amount of sample, then pushes out the sample by raising the piston, and the liquid level sensor and After switching the rotary valve to remove the air layer and inject only the sample into the column, the developing solvent is injected into the column through the short-circuit channel at a position where the piston is ascended.

[0010] When the above-mentioned fixed quantity injector is used, the sample can be sucked and stored in the fixed quantity injector, and then can be sent to the column at a required pressure. The developing solvent can be sent to the column through the short-circuit channel formed in the head member without storing the liquid in the column. As described above, when the quantitative injector is used, the path of the sample and the developing solvent to the column can be shared, so that the loss of the sample is reduced, and since the sample is directly injected into the column, the diffusion of the sample can be prevented.

In the bubble removing section, a bubble remover is provided on a line for sending the effluent from the column. The bubble extractor has an inlet for the column effluent at the top of the container, an open port, and an outlet to the detector side, and causes the liquid-feeding tube to hang down to the vicinity of the container bottom through the outlet. And, a pair of upper and lower liquid level sensors are attached to the lower part of the container, and further, solenoid valves are respectively provided on the pipes on the outlet side and the pipe connected to the open port which are continuous with the liquid feeding tube,
These solenoid valves are opened and closed by the detection signal of the liquid level sensor,
If the lower liquid level sensor does not detect the liquid, the solenoid valve on the outlet side closes and the solenoid valve on the open side opens to release the air in the container, while if the upper liquid level sensor detects the liquid, the open The liquid supply is performed by closing the electromagnetic valve on the side and opening the electromagnetic valve on the outlet side. Note that the upper sensor is not always necessary.

[0012] In the above bubble remover, the inside of the container is opened until a certain amount of column effluent accumulates in the container, so that air is vented. In addition, when a certain amount of column effluent accumulates, the air vent port is closed and a sealed state is established, so the column effluent is sent from the liquid sending tube immersed in the liquid. It is possible to reliably prevent the liquid from being sent to the detector in a state where air bubbles are mixed therein.

Further, since a detector for detecting the component of the effluent is provided downstream of the bubble remover, the component of the effluent of the column is detected by the detector, whereby the component in the effluent dividing section is detected. The start and end times of collection can be detected and the chromatographic work can be automatically controlled. In addition, since a bubble extractor is provided upstream of the detector so that bubbles do not enter the effluent, the detection accuracy of the detector can be improved.

Further, in the automatic refining device, the first
Since the sample supply unit side and the developing solution mixing / sending unit side are connected to the port of the rotary valve, the sample is sent to the quantitative injector by rotating and switching the first rotary valve, and then the developing solvent is Can be sent. In addition, the sample and the developing solvent can be sequentially sent from the metering injector to the column via the second and third rotary valves.

According to a second aspect of the present invention, the sample supply unit includes a plurality of sample containers and an auxiliary sample container connected to the sample containers. 2. The automatic purifying apparatus according to claim 1, wherein the automatic purifying apparatus is provided so as to be able to communicate with one rotary valve, and further has a connection end portion detachably connected to a sample container of another apparatus.

Since the auxiliary sample container is provided as described above, the entire amount of the sample in the sample container can be injected into the column without loss. In addition, since it can be connected to a sample container of another device, it can be used as an automatic synthesizer in combination with a generator, or can be used alone.

According to a third aspect of the present invention, in the third aspect, the developing solution mixing and feeding unit includes an HPLC pump, a gran jet device, and a plurality of developing solvent storage containers, and the developing solvent can be arbitrarily controlled by the gran jet device. And the liquid can be sent to the fixed-volume injector and the column unit via the first rotary valve by the HPLC pump, and a vacuum pump is connected to the second rotary valve, and the vacuum pump is connected to the second rotary valve. 2. A structure in which a negative pressure is introduced into a fixed-quantity injector to aspirate a sample, a washing solvent and a developing solvent.
Alternatively, an automatic purification device according to claim 2 is provided.

Since the mixing of the developing solvent by the above-mentioned gran jet device is also automatically performed based on a program input in advance, a required developing solvent can be easily prepared and sent to the column. Further, the HPLC pump is R
Since it is possible to communicate with the computer by the S232C communication circuit, the liquid sending speed can be changed under any conditions. In addition, when the negative pressure of the vacuum pump is introduced into the fixed-volume injector as described above, it is possible to more reliably send the sample and the washing solvent to the fixed-volume injector, and to remove the developing liquid between the rotary valves in advance. Thus, dilution of the sample can be prevented.

According to the present invention, the third rotary valve is a six-way rotary valve.
4. An automatic purification apparatus according to claim 1, wherein a rotary valve is connected to two columns provided in the column section, and the column can be selected and used. ing.

When two columns are attached as described above, they can be used properly as required. For example,
If an open column and a closed column are attached, purification can be performed while detecting the presence or absence of a developing solvent using the open column.

Further, according to the present invention, in claim 5, the effluent dividing section is provided with two fraction collectors, one for peak division and the other for volume division, and the column effluent is selected by selecting one of the fraction collectors. An automatic refining apparatus according to any one of claims 1 to 4 is provided.

If two fraction collectors are installed as described above, any appropriate method of peak division, time division or volume division can be adopted according to the state of the column effluent, and the usability is improved.

[0023]

Embodiments of the present invention will be described below with reference to the drawings. As shown in FIG. 1, the automatic purification apparatus using the column chromatography method according to the present invention is broadly divided into a hard part and a software part, and the hard part comprises a drive unit I and a control unit II. Drive unit I
Represents a developing solution mixing / sending section A, a sample supply section B, a sample filling section C,
It comprises a column section D, an air bubble removing section E, an effluent component detecting section F, and an effluent dividing section G.

The software section includes a refining program and a data processing program, and these programs are input to the computer CPU of the control unit II as shown in FIG. The CPU includes an output unit connected to a valve and a relay, which will be described later, provided in each of the units A to G of the drive unit I.
An input unit connected to the valve position sensor and the liquid level sensor, an A / D converter connected to the voltage output unit of the detector, a pump, and an RS232C communication circuit connected to the detector are provided. With this configuration, the drive unit I is driven based on a program stored and input to the CPU of the control unit II.

Specifically, in the automatic refining apparatus of the present invention, the inside front side of the integrated box 1 shown in FIG. 3 is partitioned by a partition plate 2 to constitute the drive unit I and the control unit II, which will be described later. Each device is stored. A caster 5 is attached to the lower end of the box 1 so that it can be moved to a required place. The box 1 is a closed type. A local exhaust fan 3 is attached to a corner of a ceiling of the box 1, a duct 4 is connected to the fan 3, and the fan 3 is connected to exhaust equipment through the duct 4. In addition, a transparent plastic door is attached to the front of the box 1 so that the state of reagent supply in the box 1 can be checked from the front side, so that the experiment can be seen from the front and the occurrence of trouble can be easily recognized. I have to.

FIG. 4 shows the overall configuration of the drive unit I composed of the above-described units A to G. The configuration of each unit will be described below in order. In FIG. 4, the solenoid valves V1 to V15 for opening and closing the flow paths provided in the flow lines L1 to L15 made of Teflon tubes are indicated by ○, and the circles attached to the circles are normally closed, the blanks are normally open, and black circles. Indicates the common side.

In the developing solution mixing / sending section A, a solvent container 1 storing a solvent to be used as a developing solution for chromatography is supplied to a HPLC pump 10 via a GranJet device 11.
2A, 12B and 12C are connected. An optical sensor PS3 is provided on a flow line L1 for sending a developing liquid, which connects the discharge side of the HPLC pump 10 and the first rotary valve 13. In the developing liquid mixing / sending section A, the solvents stored in the three kinds of solvent containers 12A to 12C are mixed with a grand jet 1
1 and mixed at an arbitrary ratio in the time sequence,
The liquid is sucked into the PLC 10, a fixed amount of solvent is sent to the first rotary valve 13 through the flow line L1, and the presence or absence of the solvent is checked by the optical sensor PS3 in the flow line L1. In addition, the HPLC pump 10 is connected to RS232.
It is connected to a computer CPU by a C communication circuit to control the liquid sending speed and on / off control. In addition, the liquid sending pressure is checked to prevent trouble.

The sample supply section B is provided with two sample containers 14A and 14B for storing a sample to be purified, and the sample in the sample containers 14A and 14B is supplied via a solenoid valve V1 or V2 to a flow line for sending a sample. The liquid is sent to the first rotary valve 13 from the flow line L2. An optical sensor PS4 is also provided on the flow line L2 to check for the presence of a sample. In addition, the flow line L2 has a connection end 60 that can connect the present apparatus to another apparatus, and is connected to another apparatus equipped with a sample container (not shown) via the connection end 60, and is connected to another apparatus. The above 3 from the sample container
The sample is transferred to the first rotary valve 13 via the one-way solenoid valve V2.
The liquid can be sent to For example, the connection end portion 60 has a structure in which a joint is attached to a pipe at the end of the flow line L2 and communicates with a joint of a pipe of a mating apparatus. Further, the auxiliary sample container 1 is connected via the solenoid valves V5 and V6.
4C and 14D, the auxiliary sample container 14C is attached to the sample container 14A, and the auxiliary sample container 14D is attached to the sample container 14A.
B is connected.

Further, there is provided a cleaning liquid container 15 for storing a cleaning liquid for cleaning the sample containers 14A and 14B after the purification, and the cleaning liquid container 15 and the sample containers 14A and 14B are provided.
B is connected to the cleaning liquid supply flow line L3 provided with solenoid valves V3, V4, V5, V6, and the sample containers 14A, 14B are connected to the vacuum pump 16 via solenoid valves V7, V8. Thus, the cleaning liquid can be sucked into the sample containers 14A and 14B.

The sample filling section C includes the first rotary valve 13, the second rotary valve 17, and the third rotary valve 18, and the first and third rotary valves 13, 18 are 6
The second rotary valve 17 is a four-way rotary valve. The first to third rotary valves 13, 17, and 18 include a position sensor P.
S1, PS2 and PS14 are installed to detect the rotational position of the rotary valve.

As shown in FIG. 5, the first rotary valve 1
Of the six ports of 3, the flow line L is connected to port 13a.
2, the flow line L1 is connected to the port 13b, and one end of the flow line L4 is connected to the port 13c. Further, one end of the flow line L16 is connected to the port 13d, and the remaining ports 13e and 13f are plugged.

The flow line L4 is provided with an optical sensor PS5.
Inlet 20 opened at the upper end surface of metering injector 20 through
a. An outlet 20b is opened at the upper end surface of the metering injector 20, one end of a flow line L5 is connected to the outlet 20b, and an optical sensor PS6 is provided.
The other end is connected to the port 17 a of the second rotary valve 17.

Port 17b of second rotary valve 17
Is connected to the vacuum pump 16 via the flow line L6, and the trap 21
An electromagnetic valve V9 is provided. The port 17c is connected to the port 18a of the third rotary valve 18 via the flow line L7. Further, the port 17d of the second rotary valve 17 is connected to the port 13d of the first rotary valve 13 via the flow line L16. Further, the port 18b of the third rotary valve 18
Is connected to the inflow port 25a of the closed column 25, and the port 18c is connected to the outflow port 25b of the closed column 25. The port 18d of the third rotary valve 18 is connected to the outlet port 26a of the open column 26 via the flow line 17, and the port 18e is connected to a flow line L8 for sending column effluent.
Is connected to

In the sample filling section C, the entire amount of the sample is introduced into the metering injector 20 from the first rotary valve 13 and stored, and then the flow line L5, the second rotary valve 17,
The sample is introduced into the closed column 25 through the flow line L7 and the third rotary valve 18. After the sample is introduced into the column, the developing solvent is sent to the column 25 through the first rotary valve 13, the flow line L4, the quantitative injector 20, the flow line L5, the second rotary valve 17, the flow line L7, and the third rotary valve 18. Liquid. At this time, the developing solvent does not accumulate in the syringe-like container 30 of the metering injector 20 and the inlet 20a and the outlet 2
0b directly through a continuous short-circuit channel.

After filling the column 25 with the sample, the first rotary valve 13 and the second rotary valve 17 are switched again, and the developing solution is directly sent to the column 25 through the flow line L16.

FIG. 6 shows the structure of the fixed quantity injector 20.
(A) and (B) hold the upper outer peripheral portion of the vertical syringe 30 with the chuck 31, and attach the chuck 3 to the bracket 33 protruding from the outer wall of the drive box 32 installed on the side.
1 is fixed and the syringe 30 is held. A piston 34 is slidably fitted in the syringe 30 and a lower end of a piston rod 35 connected to the piston 34 is attached to the substrate 3.
6 and the substrate 36 is connected to a lifting member 37 in the drive box 32.

In the drive box 32, a rod 39 is erected from a base 38 so as to be rotatable, and a screw 39a is formed on the rod 39 on the outer peripheral surface within a required range from the lower end.
A female screw hole 37a provided in the elevating member 37 is screwed into 9a. The upper end of the rod 39 is connected to a pulse motor 40, and the rod 39 is rotated by the pulse motor 40 to raise and lower the screwed lifting member 37,
The piston 34 is moved up and down in the syringe 30. The pulse motor 40 is mounted on a base 41 fixed to a side wall of the drive box 32. Further, the upper and lower bases 41 and 38
As shown in FIG. 6B, the four guide rails 4
4 and a through hole 37b through which the guide rail 44 is inserted. The guide rails 44 passing through the through holes 37b are positioned so as to surround the rod 39, so that the elevating member 37 moves up and down stably while maintaining the horizontal state, and increases the pressure resistance.

A head member 42 is removably attached to the upper end of the syringe 30. The head member 42 is provided with the inlet 20a and the outlet 20b of the upper end in the vertical direction. A groove 20c having a lower end opening communicating the lower end between the outlet 20b and the outlet 20b is provided. When the piston 34 reaches the upper limit and the upper surface of the piston contacts the lower surface of the head member 42, the groove 20c closes the lower end opening and forms a short-circuit channel 20d communicating the inflow port 20a and the outflow port 20b. ing.

That is, as shown in FIG. 7A, when the piston 34 descends, the inflow port 20a and the outflow port 20b communicate with the inside of the syringe 30, and the liquid flowing from the inflow port 20a is supplied to the syringe 30. It is stored inside and pushed out from the outlet 20b by the rise of the piston 34. On the other hand, FIG.
As shown in (B), at the upper limit of the piston 34, the inflow port 20a and the outflow port 20b are cut off from the inside of the syringe 30 to form a short-circuit channel 20d which directly communicates with the inside of the groove 20c. The liquid introduced from 20a is sent from the outlet 20b through the groove 20c.

The upper and lower ends of the syringe 30 are provided with position sensors PS7 and PS8. Further, the pressure sensor 43 is attached to the substrate 36, and the rising speed is controlled by controlling the rotation speed of the pulse motor 40 so that the pressure does not become 1 kg / cm ² or more when the piston rod 35 is raised, and a predetermined pressure or more is applied. Sometimes, the pulse motor 40 is stopped. In this embodiment, when a pressure of 5 kg / cm ² is applied, the pressure sensor 43 outputs a detection signal to stop the pulse motor 40.

In the fixed-quantity injector 20 having the above structure, the sample is sucked from the sample container 14A or 14B through the inflow port 20a and stored in the syringe 30 while the piston 34 is lowered. After the required amount has been charged, the pulse motor 40 is rotated to raise the piston 34 so that the pressure is within 1 kg / cm ² , and the stored sample is sent to the second rotary valve 17 through the outlet 20b. When the piston 34 reaches the upper limit, the head member 42
Abuts the lower surface of the syringe 30 and the inside of the syringe 30 and the inflow port 20a,
While the outlet 20b is cut off, the short-circuit channel 20d connects the inlet 20a and the outlet 20b through the groove 20c.
Is formed. In this state, the developing solvent introduced from the inflow port 20a side flows through the short-circuit channel 20d to the flow line L5.
Sent to When the optical sensors PS5 and PS6 detect the liquid in the syringe 30 when the piston 34 rises, the two rotary valves are switched to inject the liquid into the column.

The column section D is the closed column 25
And an open column 26. The third rotary valve 18 in six directions is switched so that the column 25 or the column 26 can be selected and used. open·
In the column 26, a glass rod type optical liquid level meter 28 is provided in a container 27.
Is attached, and the presence or absence of the developing solvent is checked.

In the bubble removing portion E, the third rotary valve 1
A bubble extractor 50 for column chromatography shown in FIG. 8 is provided in a flow line L8 for sending column effluent, which is connected to the port 18e of No. 8. The bubble extractor 50 detects bubbles in the liquid flowing out of the column 25, excludes the detected bubbles, and passes only the effluent through the flow line L9 to the detector 51 for detecting the effluent component in the detection unit F. The liquid is sent.

The bubble eliminator 50 includes a glass container 52 having a conical portion continuous with a lower portion of a cylindrical portion, and a connecting pipe portion 52a protruding from one side of an upper peripheral wall of the glass container 52 is connected to a flow line. While connected to L8, a connection pipe portion 52b protruding from the other side is connected to an open line L10 for removing air bubbles, and an electromagnetic valve V10 is provided in the open line L10. In addition, a cap 5 to cover the upper end of the glass container 52.
3 is provided with a through hole 53a at the center thereof, a Teflon tube 54 is suspended from the upper end into the glass container 52 through the through hole 53a, and a lower end opening 54a of the Teflon tube 54 is provided.
Is opened at the center of the bottom surface protruding downward of the glass container 52. The upper end of the Teflon tube 54 is a flow line L9.
And the detector 51 and the solenoid valve V are connected to the flow line L9.
12 are interposed.

A pair of upper and lower liquid level sensors 55A and 55B are attached to the lower outer peripheral portion of the glass container 52.
The pipe 54 projects downward from the lower liquid level sensor 55B.

In the bubble eliminator 50 having the above configuration, the column 25
The liquid that has flowed out through the third rotary valve 18 and the flow line L8 flows into the glass container 52 from the upper connecting pipe portion 52a along the wall surface, and accumulates in the container 52. At this time, the solenoid valve V10 is open and V12 is closed. When liquid is detected by the upper liquid level sensor 55A, the electromagnetic valve V10 is closed and the electromagnetic valve V12 is opened, and the liquid in the container 52 is led out to the flow line L9 by the Teflon tube 54 by liquid pressure and air pressure. When the lower liquid level sensor 55B stops detecting the liquid due to the derivation of the liquid, the electromagnetic valve V10 is opened and the electromagnetic valve V12 is closed, and the liquid is stored in the container 52 until the upper liquid level sensor 55A detects the liquid level. Save. In this manner, the glass container is opened to release air until a certain amount or more of liquid is collected inside the glass container 52, and when a certain amount of liquid is collected, the tube 5 immersed in the liquid is removed.
Since the configuration is such that the liquid is led out from 4, the air bubbles can be reliably prevented from being introduced into the flow line L9.

The bubble remover is not limited to the above embodiment, but may be a lower level sensor alone without an upper level sensor. However, in this case, the liquid level stops around the level unless the liquid sending amount is equal to or more than a certain value, so that the ON / OFF repetition increases.

The effluent component detector 51 provided in the flow line L9 detects the components of the effluent with visible light, ultraviolet light, or the like, and outputs the signal to the computer in proportion to the intensity of the absorbance. And In this embodiment, UV
A detector is used. The detector 51 is connected by an RS232C communication circuit, and controls the setting of a detection wavelength, ON / OFF of a UV lamp, and the like.

The effluent dividing section G has a large fraction collector 55 and a small fraction collector 56,
Large fraction collector 55 is flow line L12
Is connected to the UV detector 51 via the. Solenoid valves V12, V13, and V14 are interposed in the flow line L12, and the small fraction collector 56 is connected to V14 through the flow line L13.

The large-sized fraction collector 55 has a plurality of large-capacity test tubes 55a mounted on a base 55b.
5a is sequentially filled and collected. On the other hand, the small fraction collector 56 has a large number of small-volume test tubes 56a mounted on a base 56b. When the effluent is divided by time or weight, the effluent is sequentially filled into the test tubes 56a and collected. ing.

If no peak is observed in the effluent,
The liquid is led to the drain tank 59 through the flow line L14 connected to the solenoid valve V13. A glass rod type optical liquid level sensor 61 is also attached to the drain tank 59, and a flow line L14, a solenoid valve V15
Through the waste liquid trap 21.

The above-mentioned solenoid valves V1 to V15, rotary valves 13, 17, 18, vacuum pump 16, HP
In the control unit II for controlling the driving of the LC pump 10 and the motor 40 of the fixed-quantity injector 20 and the like, a notebook personal computer housed in the box 1 is used as the computer CPU to achieve compactness. In addition, an I / O expansion unit is added for transmitting and receiving signals between the computer CPU and the drive unit I, and a parallel input / output module, an A / D conversion module, and an RS232C communication circuit are provided.

The program to be input to the computer CPU includes a column selection, a time, a test tube number of a fraction collector, a flow rate of a developing solution, a grand jet condition, a detection wavelength, an experimental code number, etc., in an interactive manner with a monitor screen. The refiner can be operated by inputting it.

The program is set so as to have the functions listed below. Before starting the apparatus, the presence / absence of a solvent, detection of the initial setting position of the fixed quantity injector 20, the rotary valves 13, 1
It also has a self-diagnosis function for detecting the rotational positions of 7 and 18 so that it can be operated only in a normal initial state.

The presence or absence of a solvent in the chromatography, the liquid sending pressure,
The tester constantly checks the fraction collector for remaining test tubes, and if there is any abnormality, suspends chromatography and waits for instructions from the experimenter. When the chromatography is continued according to the instruction of the experimenter, the chromatography can be resumed from the interrupted time. When the chromatography is stopped and terminated, the data of the time up to that point is registered in the hard disk so that the next operation such as washing can be performed.

Detector 5 using RS232C communication circuit
1. Since the HPLC pump 10 is controlled, the liquid sending speed and the detection wavelength can be changed even during chromatography.

When the fraction division method is peak division, the output of the detector 51 is constantly monitored and its state is displayed in a graph. When an output and a slope exceeding a certain level are detected, the effluent is discharged from the drain tank 59 to the fraction collector 5.
The solenoid valve is switched to 5 and divided. Monitor the slope even during splitting, even if the peak contains multiple peaks,
The test tube is switched at the position of the valley of each peak so that it can be divided for each peak. In addition, when the test tube of the fraction collector is switched, a line is displayed on the graph on the monitor screen so that the test tube number and the graph can correspond accurately. When the effluent is collected by the fraction collector 56 by dividing the time (volume), the effluent is monitored by the detector 51 and displayed in a graph as in the case of the peak division. When a test tube is switched by the fraction collector 56, the output signal is displayed as a line on a graph on a monitor screen so that the number of the test tube and the graph can correspond exactly. All graph data displayed on the monitor is recorded on the hard disk.

Next, a refining process performed by driving the drive unit I of the refining apparatus based on the program shown in FIG. 9 input to the computer CPU will be described.

First, the column to be used and the solvent containers 12A to 12C storing the developing solvent are set. Then, the manual pump is pushed to move the HPLC pump 10, and it is confirmed that the developing solvent is normally introduced into the column. When the purifier is used alone, a sample is put in the sample container 14A and / or 14B using a pipette or the like. Next, the cleaning liquid is put in the auxiliary containers 14C and 14D. In addition, when used by being attached to another device, a tube connected to a sample container of another device is connected to the solenoid valve V2.

After the necessary parts are connected, the refining conditions in step # 1 are input to the computer CPU. That is, in the computer, the compound name, code number, sample container selection, column selection, purification time, fraction collector selection, fraction collector test tube number,
The flow rate of the developing solvent, the wavelength of the detector, etc. are input.

After inputting the purification conditions and before starting the purification, the initial state of step # 2 is confirmed. That is, FIG.
, The output signals PS 1, 2, 14 of the rotary valves 13, 17, 18 indicate that the rotary valves 13, 17, 18
Position of the rotor, the fixed quantity injector 2 by the optical sensors PS7 and 8
The position of the piston 34 and the presence or absence of the solvent are checked by the optical sensors PS3, 5, and 6.

If there is no abnormality in checking the initial state in FIG.
In step # 3, the sample is transferred from the sample container to the quantitative injector 20. That is, first, the HPLC pump 10 is stopped,
The motor 40 of the metering injector 20 is driven, the piston 34 is pulled down via the piston rod 35, the solenoid valve V1 is opened, and a sample is introduced from the sample container 14A into the metering injector 20 via the first rotary valve 13. . Quantitative injector 2
At 0, the sample is introduced into the syringe 30 from the inlet 20a.

After confirming the transfer of the sample by the optical sensor PS4 of the flow line L2 and the optical sensor PS5 of the flow line L4, the lowering of the piston 34 is stopped after a predetermined time. If transfer of sample cannot be confirmed or piston 3
When 4 is too low and is detected by the lower position sensor PS8, the fact is displayed on the monitor and the operation is stopped.

Next, the vacuum pump 16 and the solenoid valve V
8, V6 is turned on, and the sample container 1 is
Transfer the cleaning solution to 4A. Next, the solenoid valves V8 and V6 are turned off and V9 is opened, and a negative pressure is introduced into the fixed quantity injector 20, so that the sample remaining in the sample container 14A, the flow line L2, the first rotary valve 13, and the flow line L4 is obtained. Is transferred to the metering injector 20 while washing. After confirming that there is no sample in the line with the optical sensors PS4 and PS5, the vacuum pump 16 is stopped and the solenoid valve V
Close 9. This prevents sample loss in the line,
All samples can be transferred into the syringe 30.

Next, the air in the fixed quantity injector 20 is removed in step # 4. That is, after opening the solenoid valve V9, the motor 40 is driven to raise the piston 34. As the piston 34 rises, the air above the syringe 30 becomes
The gas is exhausted from the outlet 20b through the flow line L5, the second rotary valve 17, the vacuum line L6, and the solenoid valve V9, and is discharged from the inlet 20a.
4. The first rotary valve 13, the flow line L2, and the valves V1 and V2 are also exhausted.

When air is removed from the fixed quantity injector 20 and the optical sensors PS5 and PS6 disposed above the inflow port 20a and the outflow port 20b detect the liquid level of the sample, step #
The sample in the quantitative injector 20 of No. 5 is packed in the column 25.

Specifically, first, the first rotary valve 13 and the second rotary valve 17 are rotated to switch the line as shown in FIGS. 10 (A) and 10 (B). That is, while the flow line L4 and the flow line L2 are cut off, the flow line L5 and the flow line L7 are connected continuously, and the sample drawn out from the quantitative injection device 20 flows through the flow line L
5, a line is connected so that liquid can be sent to the column 25 through the second rotary valve 17, the flow line L7, and the third rotary valve 18. At this time, the florins L4 and L1 also communicate with each other, but there is no flow of the liquid from L4 to L1 due to the check valve of the HPLC pump 10.

Further, the motor 34 is driven by the metering injector 20 to raise the piston 34, and the sample is filled in the column 25 through the above-mentioned line. The pressure at the time of filling is detected by the pressure sensor 43, and the drive of the motor 40 is controlled by a computer so that the pressure does not exceed 1 kg / cm ² .
If the pressure becomes 5 kg / cm ² on the way, this is displayed on the monitor screen and stopped.

When the piston 34 reaches the upper end of the syringe 30, the position sensor PS7 detects it, and the HPLC pump 1
0, the grand jet 11 is driven, and the developing solvent is sent to the metering injector 20 via the flow line L1, the first rotary valve 13, and the flow line L4.

In the fixed quantity injector 20, the piston 34 at the upper limit position contacts the lower end surface of the head member 42 and
c, the lower end opening is closed, and the groove 20c forms a short-circuit channel 20d that connects the inflow port 20a and the outflow port 20b. Therefore, the developing solvent sent to the inflow port 20a flows out of the outflow port 20b through the communication path. That is, the metering injector 20
Is used as a flow path for the developing solvent.

As described above, the developing solvent is sent to the column 25 filled with the sample, so that step # 6 is performed.
Chromatography is started. When the samples in the flow lines L4 and L5 and the short-circuit channel 20d of the quantitative injector 20 are completely washed out with the developing solvent and filled in the column 25, the syringe 30
In order to prevent the internal pressure from being applied, the first rotary valve 13 and the second rotary valve 17 are switched, and the developing solvent is directly sent from the flow line L4 to the column 25 as shown in FIG.

The effluent from the column 25 is sent to the flow line L8 via the third rotary valve 18, and the bubbles are reliably removed by the above-described operation by the air bubble remover 50 provided in the flow line L8. The liquid is sent from the air bubble remover 50 to the flow line L9, and the components are detected by the UV detector 51.

When the fraction division method is the peak division, the output of the detector 51 is constantly monitored and its state is displayed as a graph. When the output (peak) and the slope exceeding a certain level are detected, the solenoid valve V13 And the effluent is sent from the drain tank 59 to the fraction collector 55, and is sequentially collected in the test tube 55a. Detector 5
If no peak is detected at 1, the division is terminated.

The same applies to the case where the fraction is divided by time (capacity). The solenoid valve V14 is switched to send the effluent to the fraction collector 56, and the test tube 56a
To collect sequentially.

When steps # 7 to # 13 are completed, the data is registered in step # 14, and the HPLC pump 1 is used to complete the chromatography in step # 15.
0, the detector 51 and the Granjot 11 are stopped, and the required solenoid valve is switched.

Next, the sample container 14A in step # 16
Is washed. That is, the vacuum pump 16 is driven, and the electromagnetic valves V3 and V8 are turned on to turn the cleaning liquid container 1 on.
From 5 the washing liquid is introduced into the sample container 14A. Next, the solenoid valves V3 and V8 are sequentially turned off, and then the solenoid valves V9 and V1 are turned on to wash the cleaning liquid in the sample container 14A via the first rotary valve 13 and the second rotary valve 17 into the syringe of the fixed quantity injector 20. Discard while drying the inside of 30 and dry.

After purifying the sample stored in the sample container 14A, the sample stored in the sample container 14B can be subsequently purified by the same steps as described above. Further, the sample stored in the sample container of another device can be purified in the same process as described above by connecting the connection end portion 60 to another device.

[0078]

As is clear from the above description, in the automatic purifying apparatus for purifying a sample by the column chromatography method according to the present invention, since the quantitative injection device is used, the sample from the sample container to the column is removed. And the transfer of the developing solvent from the container storing the developing solvent to the column,
This can be done via one metered dose injector. As a result, the liquid transfer path from the sample container and the developing solvent container to the quantitative injector can be partially shared, and the liquid transport path from the quantitative injector to the column can be shared, so that there is no loss of the sample. Further, unlike the loop method, the sample is directly injected into the column, so that the diffusion of the sample can be prevented. Therefore, the separation ability in the column is improved. Further, a larger amount of sample can be injected into the column than in the loop method.

Further, since a bubble eliminator for the column effluent is provided, bubbles mixed into the column effluent can be reliably removed, and the detection accuracy of a detector installed downstream can be improved. .

Further, the automatic purifier is connected and combined with a sample supply section, a developing solution mixing / sending section, a sample filling section, a column section, a detecting section, an effluent splitting section, etc. by a reasonably designed liquid sending path. In addition, when the control unit performs the refining process by fully automatic control, it is conventionally possible to save labor and time for the refining work that requires a long time constraint. Also, because the required work is performed automatically, regardless of the skill level of the experimenter,
Achieve uniform results.

The automatic purifying apparatus of the present invention can be detachably connected to another apparatus to purify the sample stored in the sample container of the connected apparatus. As described above, the present purification apparatus can be used alone, and it can also be used in combination with a production reaction apparatus to be integrated and used as an automatic synthesizer, and can be used properly according to the application.

Further, in the present invention, the necessary purification work is performed only by inputting the purification conditions into the computer. If the purification conditions are changed and input, the purification work in a different mode is performed, and the change of the purification conditions can be easily performed. Can be performed.

[Brief description of the drawings]

FIG. 1 is a block diagram of an entire purification apparatus according to the present invention.

FIG. 2 is a block diagram showing a relationship between a control unit and a drive unit of the purification device.

FIG. 3 is an overall schematic view of the purification device.

FIG. 4 is a drawing showing the overall configuration of the above-mentioned purification device.

FIG. 5 is an enlarged view of a main part of FIG. 4;

FIG. 6A is a cross-sectional view of a syringe-type column injector used in the purification device, and FIG. 6B is a perspective view of a main part.

FIGS. 7A and 7B are explanatory views showing flow paths that change depending on the position of a piston of the syringe type column injector.

FIG. 8 is a front view of an air bubble remover used in the purification device.

FIG. 9 is a flowchart showing a purification process of the purification device.

FIGS. 10A and 10B are schematic diagrams showing a switching state of a rotary valve.

[Explanation of symbols]

 1 Box 10 HPLC Pump 12A to 12C Developing Solvent Container 13 First Rotary Valve 14A, 14B Sample Container 16 Vacuum Pump 17 Second Rotary Valve 18 Third Rotary Valve 20 Syringe Type Column Injector 20a Inlet 20b Outlet 20c Groove 20d Short Circuit Channel 25, 26 Column 30 Syringe 34 Piston 42 Head member 50 Bubble extractor 51 Detector 55, 56 Fraction collector

Claims (5)

[Claims] 1. A driving unit comprising: a driving unit; and a control unit, wherein the driving unit comprises:
It comprises a sample filling section, a column section, a bubble removing section, an effluent component detection section, and an effluent splitting section. The developing liquid mixing / sending section and the sample supply section are each provided via a flow line provided with an electromagnetic valve. Connected to a first rotary valve, the first rotary valve being connected via a flow line to a metering injector provided at the sample filling section, and the sample metering section being connected to the metering injector via a flow line at the sample filling section. 2,
The third rotary valve is connected to the third rotary valve in sequence, the third rotary valve is connected to the column, and the developing solution is sent to the column after the sample is sent to the column from the quantitative injector. Effluent from the effluent is sent to the effluent splitting section through a flow line provided with the bubble bleeding section and the effluent component detecting section, and the electromagnetic valve and the rotary valve are supplied by a signal from the control unit. An automatic refining apparatus characterized in that the operation of a valve is controlled. 2. The sample supply section includes a plurality of sample containers and an auxiliary sample container connected to the sample containers, selects the plurality of sample containers and communicates with the first rotary valve, 2. The automatic purifying apparatus according to claim 1, further comprising a connection end portion to which a sample container of another apparatus is detachably connected to communicate with the first rotary valve. 3. The developing liquid mixing / sending unit includes an HPLC pump, a gran jet device, and a plurality of storage containers for developing solvents. In this configuration, the liquid can be sent to the metering injector and the column section via the first rotary valve, and a vacuum pump is connected to the second rotary valve, and the negative pressure of the vacuum pump is introduced into the metering injector. 3. The automatic purifying apparatus according to claim 1, wherein the sample, the washing solvent and the developing solvent can be sucked. 4. The third rotary valve is composed of a six-way rotary valve, and the third rotary valve is connected to two columns provided in the column section so that a column can be selected and used. Claims 1 to 3
The automatic refining device according to any one of the above. 5. The effluent dividing section is provided with two fraction collectors, one for peak division and one for volume division, and selects one of the fraction collectors to divide the column effluent. Item 5. The automatic refining device according to any one of Items 4.