JP3854716B2 - Liquid chromatograph - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a liquid chromatograph, and more particularly to a liquid chromatograph that uses a plurality of columns to regenerate (wash and re-stabilize) the other column during analysis in one column.
[0002]
[Prior art]
In a conventional liquid chromatograph, there is a method of regenerating (washing and re-stabilizing) the other column during analysis in one column using an analysis pump, a regeneration pump, a valve, and a plurality of columns. In a liquid chromatograph with such a configuration, the target operation (analysis, cleaning, re-stabilization) of each pump is disassembled, a time chart is created for the pump operation and valve operation, and each pump Adjustments were made in consideration of the consistency of the time chart, and the entire gradient program had to be created.
[0003]
[Problems to be solved by the invention]
In the above-described prior art, since the creation of the gradient program in consideration of the time charts of the individual pumps and valves has been performed manually by the operator, the creation of the program has been performed by experienced technicians. Therefore, there is a problem that operators who can create a program are limited. Another problem is that mistakes are likely to occur when developing a program such as a plurality of pumps, and the analysis cannot be started without confirmation several times in advance. An object of the present invention is to provide a liquid chromatograph that automatically creates a program for a plurality of pumps and valves by inputting a series of operations (analysis, washing, re-stabilization) in a single column.
[0004]
[Means for Solving the Problems]
In the above object, the present invention is characterized in that a first pump for feeding an eluent for analysis, a second pump for feeding an eluent for column regeneration and re-stabilization, and injecting a sample to be analyzed. A sample injection unit, at least two columns provided in parallel to the first pump for separating the sample for each component, a detection unit for detecting the sample separated by the column, and the devices. A liquid chromatograph configured to control an eluent having a different composition from the second pump to the other column during sample analysis in one of the columns; A gradient program creating means for creating a gradient program including the second pump from the set gradient program for the first pump;
[0005]
DETAILED DESCRIPTION OF THE INVENTION
Hereinafter, embodiments of the present invention will be described using examples.
[0006]
FIG. 1 shows the configuration of a first embodiment of the liquid chromatograph of the present invention. The pump 1 pressurizes the liquid while changing the composition ratio of the eluent 3 and the eluent 4 by switching the proportioning valve 2 and feeds the liquid at a constant flow rate. The sample injection device 5 has a sample injection port and a high-pressure channel switching valve for sample injection, and injects the sample into the channel. The injected sample is separated into single components by the column 19 and eluted in order from components having a small interaction with the column filler. Each component in the eluate is detected as a peak by a detector 9 having a plurality of detection channels using ultraviolet light as a light source. The condition of each unit is set by the controller 13.
[0007]
A configuration example of the controller 13 is shown in FIG. The controller 13 includes an arithmetic device 14, a display device 15, an input device 16, a storage device 17, and an input / output device 18, and a pump / sample injection device and a valve are controlled by contact control or communication control via the input / output device 18. Is controlling.
[0008]
FIG. 3 is an example of a gradient program creation initial setting screen for changing the eluent composition in the controller 13. In this embodiment, the controller 13 uses a personal computer.
[0009]
Next, an example of creating a gradient program in this embodiment will be shown.
[0010]
First, the time for performing the gradient, the time for washing and re-stabilizing the column, and the flow rate are requested as “Gradient Time”, “Regeneration Time”, and “Flow Rate”, respectively. “Deley Volume (flow path stabilization capacity)” based on the volume of the mixer that mixes the solvent indicates the value input in advance at the time of system configuration, and does not need to be input here unless particularly required.
[0011]
Here, as an example, if 6 minutes is entered in “Gradient Time” and 6 minutes is entered in “Regeneration Time” and the OK button is pressed, the screen of FIG. 4 is displayed. In this example, as an initial display, the composition of the eluent B at the start of the gradient and the column stabilization is 20%, the composition of the eluent B at the end of the gradient is 80%, and the composition of the eluent B at the time of column washing is The composition is defined as 100%, and the column washing time is defined as 1/2 of the input regeneration time (RegenerationTime).
[0012]
The right side of the screen shown in FIG. 4 shows the set gradient program as a table of time and A / B eluent composition. The left side of the screen shows the composition ratio of the eluent B with a graph with respect to the time axis. With the graphical display of the gradient program on the left side of the screen, it is possible to visually confirm what kind of gradient program is created, and it is possible to easily confirm whether or not the target program is created.
[0013]
Hereinafter, a method of creating a gradient program in the controller 13 will be described with reference to a screen example.
[0014]
FIG. 5 shows an example in which the setting of the eluent composition ratio at time 0 is changed. When a point that intersects with time 0 in the graph showing the composition ratio of the eluent is clicked, a point on the clicked graph is selected, and a square mark confirming the selection is entered. The initial eluent composition ratio of the gradient program can be set by dragging this point with the mouse. If the point at time 0 is selected, this point moves only on the composition ratio axis of the eluent B. The current pointer position is displayed at the bottom of the screen as the cursor position. FIG. 6 is a screen example when the point indicating the eluent composition at time 0 is dropped at 40%. In accordance with the position of the pointer, the row of time 0 of the gradient program display in the table on the right side of the screen is also changed to the same numerical value, that is, “40” for the B liquid composition ratio. Similarly, the composition ratio of the B liquid during column stabilization is also changed to “40”.
[0015]
Similarly, the initial eluent composition at the end of the gradient can also be changed from the graph. When the change is made, the numerical value of the corresponding time in the gradient program display in the table on the right side of the screen is changed.
[0016]
FIG. 7 shows an example of setting the eluent composition during column washing. When the cleaning portion in the graph display is clicked, the cleaning portion line (in the example, the 100% B portion line) is selected, and a thick line indicating that it has been selected is displayed. The composition ratio of the B liquid at the time of washing | cleaning can be changed by dragging this line. This line moves perpendicular to the time axis by dragging. FIG. 8 shows an example in which the liquid B is dropped at a point where the composition ratio is 90%. In accordance with the change in the composition ratio of the B liquid from the graph to 90%, the composition ratio of the B liquid is changed to “90” in the rows of the time 6.1 and 9.0 in the gradient program display according to the table on the right side of the screen. Be changed.
[0017]
FIG. 9 shows an example in which the ratio between the column washing time and the re-stabilization time is changed. When a boundary line (vertical line at 9 minutes) between the cleaning part and the re-stabilization part in the graph display is clicked, the boundary line is selected, and a bold line display indicating the selection is made. By dragging this line, the ratio between the column washing time and the re-stabilization time can be changed. This line moves horizontally with the time axis by dragging. FIG. 10 shows an example in which the boundary line is dropped in 8 minutes. Along with the change of the boundary line from the graph to 8 minutes, the time “9.0” and “9.1” of the gradient program display by the table on the right side of the screen is changed to “8.0” and “8.1”. Changed to
[0018]
In the above example, the gradient program is changed using a graph. However, the table on the right side of the screen is not necessarily displayed at this time, and the program can be changed even when the table is not displayed.
[0019]
FIG. 11 shows an example in which the gradient program setting is changed by displaying the gradient program using a table. Fig. 11 shows an example of changing the eluent composition at the time 0.0 minute line, that is, the gradient start point. Click the column displaying the composition of eluent B in the 0.0 minute line in the table. Then, the column is highlighted and a numerical value can be input. When “40” is input as the composition of the B eluent at 0.0 minutes using the input device 16, the screen is changed as shown in FIG. That is, “60” obtained by subtracting 40 from 100 is automatically displayed in the column of the composition ratio of the eluent A, and the graphical display on the left side of the screen is also changed to a position of 40% so as to match the input from the table. At this time, since the 0.0 minute line is the gradient start point, the composition ratio of re-stabilization is also changed to 40%, which is the same as the composition ratio of the gradient start point.
[0020]
In the present embodiment, as described above, since the gradient program can be created and edited on the graphical display screen, the program for setting the target eluent composition change can be recognized sensibly and easily. There is an effect that can be created. In addition, when creating and editing gradients using tables, the graphical display of eluent composition changes changes simultaneously, which also has the effect of preventing setting errors.
[0021]
FIG. 13 shows the configuration of the second embodiment of the liquid chromatograph of the present invention. In this embodiment, a plurality of columns are provided for one analysis, and the gradient program creation in the case of such a configuration is shown.
[0022]
In the configuration of FIG. 13, the two analytical pumps 1 supply the liquid at a constant flow rate by pressurizing the liquid while changing the composition ratio of the eluent 3 and the eluent 4 by switching the proportioning valve 2. The sample injection device 5 has a plurality of sample injection ports and a high-pressure channel switching valve for sample injection, and injects different samples into the plurality of channels. The injected sample is guided to the ten-way valve 6. The ten-way valve 6 is currently set in a flow path indicated by a solid line, and the sample is separated into single components by the column A of 7 and eluted in the order of components having a small interaction with the column packing material. Each component in the eluate is detected as a peak by a detector 9 having a plurality of detection flow paths using ultraviolet light as a light source through the ten-way valve 6 again. Each eluate flow path is connected to the fraction collector 10, and each component is collected in a container such as a test tube that is different for each component. During this analysis, the regeneration pump 11 changes the composition ratio of the eluent 3 and the eluent 4 by switching the proportioning valve 2, pressurizes the cleaning liquid and subsequently the re-stable liquid, and sends it at a constant flow rate. Liquid. The washing solution and the re-stabilizing solution are branched into two, and two column Bs of 8 are simultaneously washed and re-stabilized through two ten-way valves 6. The cleaning liquid and the re-stabilization liquid are collected in the drain bottle 12.
[0023]
FIG. 14 shows the vicinity of the ten-way valve 6 in detail. When the ten-way valve 6 is set to a flow path indicated by a solid line, the analysis pump 1 is communicated with the column A of 7 and the regeneration pump 11 is communicated with the column B of 7. At this time, the sample is analyzed in the column A of 7, and the eluate is guided to the detector 9. The column B of 8 is washed and re-stabilized, and the eluate is guided to the drain bottle 12. When the ten-way valve 6 is switched to the dotted line side, analysis is performed on the column B of 8, and washing and re-stabilization of the column A of 7 are performed.
[0024]
The analysis pump 1, the regeneration pump 11, the sample injection device 5, and the ten-way valve 6 are controlled by a controller 13. The configuration of the controller 13 is shown in FIG. 3 as in the first embodiment. The controller 13 includes an arithmetic device 14, a display device 15, an input device 16, a storage device 17, and an input / output device 18, and a pump / sample injection device and a valve are controlled by contact control or communication control via the input / output device 18. Is controlling.
[0025]
FIG. 15 shows a gradient program created by the controller 13 in the same manner as in the first embodiment in the second embodiment. FIG. 18 shows the program displayed in a table format. In this example, the analysis time is set to 8 minutes, the washing time is set to 2 minutes, the re-stabilization time is set to 5 minutes, and the flow rate is set to 2 mL / minute. Further, the channel stabilization capacity mainly resulting from the mixer capacity for mixing the solvent installed in the channel is 4 mL in the system of this example.
[0026]
Since the controller 13 of the present embodiment has two columns for analysis and regeneration, as shown in FIG. 17, from the gradient program set as described above, A: analysis time, B : Washing time, C: Re-stabilization time, D: Flow rate, E: Flow stabilization capacity parameters are obtained, and each calculation is performed based on these parameters, and the conditions of the analysis pump and regeneration pump are automatically determined. Perform the calculation. In this example, A = 8 minutes, B = 2 minutes, C = 5 minutes, D = 2 mL / min, and E = 4 mL. Therefore, the shortest cycle of the analytical pump is 10 minutes by the calculation method shown in FIG. The shortest cycle of the regeneration pump is calculated as 7 minutes, and the sample injection cycle is 10 minutes. From this, the flow path stabilization time of the analysis time of the analysis pump is calculated as 2 minutes, and the re-stabilization time of the regeneration pump is calculated as 8 minutes. As described above, the analysis pump / regeneration pump and ten-way valve program created according to the conditions calculated in this example is as shown in FIG. This is shown in a table format in FIG.
[0027]
Using the chromatograph of this example, in a system having two columns for analysis and regeneration, a program for the analysis pump, regeneration pump and valve can be created simply by inputting a normal gradient program. Therefore, there is an effect that the throughput can be easily improved by washing and re-stabilizing the other column during the analysis in one column.
[0028]
【The invention's effect】
According to the present invention, in a system having a plurality of columns and two columns for analysis and regeneration in each system, an analysis pump, a regeneration pump and a valve can be obtained only by inputting a normal gradient program. Therefore, there is an effect that it is possible to easily improve the throughput by washing and re-stabilizing the other column during the analysis in one column.
[Brief description of the drawings]
FIG. 1 is a configuration diagram of a first embodiment of the present invention.
FIG. 2 is a system controller configuration diagram of the present invention.
FIG. 3 is an example of an initial input screen according to the present invention.
FIG. 4 is an example of a gradient program initial creation screen of the present invention.
FIG. 5 is an example of a gradient program creation screen of the present invention.
FIG. 6 is an example of a gradient program creation screen of the present invention.
FIG. 7 is an example of a gradient program creation screen of the present invention.
FIG. 8 is an example of a gradient program creation screen of the present invention.
FIG. 9 is an example of a gradient program creation screen of the present invention.
FIG. 10 is an example of a gradient program creation screen of the present invention.
FIG. 11 is an example of a gradient program creation screen of the present invention.
FIG. 12 is an example of a gradient program creation screen of the present invention.
FIG. 13 is a flow path configuration diagram of a second embodiment of the present invention.
FIG. 14 is a detailed view of the vicinity of a ten-way valve according to a second embodiment of the present invention.
FIG. 15 is a gradient program set in the second embodiment of the present invention.
FIG. 16 is a time program automatically created in the second embodiment of the present invention.
FIG. 17 is an automatic program creation method according to the second embodiment of the present invention;
FIG. 18 is a gradient program set in the second embodiment of the present invention.
FIG. 19 is a time program automatically created in the second embodiment of the present invention.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Analytical pump, 2 ... Proportioning valve, 3 ... Eluent A, 4 ... Eluent B, 5 ... Sample injection apparatus, 6 ... Ten way valve, 7 ... Column A, 8 ... Column B, 9 ... Detector DESCRIPTION OF SYMBOLS 10 ... Fraction collector, 11 ... Regeneration pump, 12 ... Drain bottle, 13 ... System controller, 14 ... Arithmetic unit, 15 ... Display device, 16 ... Input device, 17 ... Storage device, 18 ... I / O device, 19 ... column.

Claims (1)

A first pump for supplying an eluent for analysis, a second pump for supplying an eluent for column regeneration and re-stabilization, a sample injection part for injecting a sample to be analyzed, and the sample for each component And at least two columns provided in parallel to the first pump, a detection unit that detects a sample separated by the column, and a control unit that controls the devices, In a liquid chromatograph for supplying an eluent of another composition from the second pump to the other column during sample analysis in one of the columns,
To the control unit, to have a gradient program creating means for creating a gradient program, including the second pump from the gradient program for the first pump is set arbitrarily,
The gradient program creating means of the control unit creates a gradient program including the second pump using the analysis time, cleaning time, re-stabilization time and flow path stabilization capacity as parameters,
Sample which is the larger of the total time of analysis time and the time obtained by dividing the channel stabilization capacity by the flow rate of the first pump or the total time of washing time and re-stabilization time A liquid chromatograph characterized by having an injection cycle .

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