JP4536570B2 - Liquid chromatograph analyzer - Google Patents

Description

  The present invention relates to a liquid chromatograph analyzer, and more particularly to a liquid chromatograph analyzer that regenerates or cleans an analysis column.

  In the liquid chromatograph analyzer, the analysis column is regenerated or washed for each analysis to improve the analysis accuracy. As a method for regenerating or washing this analytical column, a gradient liquid feeding method using a gradient system for feeding an eluent containing a solvent intended for column regeneration or washing is generally used as an eluent. Has been.

  Patent Document 1 describes a cleaning method for efficiently cleaning an analytical column used for ion chromatography. The technique described in Patent Document 1 is a method of washing an analytical column with a mixed solution of pyromellitic acid and 2-amino-2-hydroxymethyl-1,3-propanediol.

JP-A-6-317572

  By the way, when the analytical column is regenerated or washed using the gradient liquid feeding method, even if the cleaning method described in Patent Document 1 is used, in addition to the analytical column regeneration time, measurement is started after returning the column to the initial state. Time (time required for conditioning) is required, and it is difficult to apply to high-throughput analysis that has been desired in recent years.

  In addition, the isocratic liquid delivery method (single liquid delivery) is ideal for routine analysis such as quality control of pharmaceuticals. For the purpose of analytical column regeneration or cleaning, as described above, separately from the pump, Therefore, there is a problem that a gradient system must be purchased for feeding the eluent, and the system becomes expensive.

  An object of the present invention is to realize a liquid chromatographic analysis method and apparatus capable of regenerating or washing an analytical column in a short time even with an isocratic liquid feeding method, while having an inexpensive configuration.

In order to achieve the above object, the present invention is configured as follows.
(1) In a liquid chromatograph analyzer, a solution feeding means for feeding a solution, a sample introduction means for introducing a sample to be measured into the solution, and a sample to be measured can be adsorbed An elution analysis column, a detection means for detecting a measurement target sample eluted from the analysis column, a solution suction / discharge means capable of sucking a washing liquid or a regenerating liquid and holding and discharging a predetermined amount; A first flow path state in which the solution fed from the liquid feeding means is supplied to the analytical column via the sample introduction means, and the solution sent from the liquid feeding means is passed through the sample introduction means. first flow path switching for switching between the second flow path state for supplying the cleaning liquid or regeneration liquid discharged from the suction and discharge means to said sample introducing means with no supply to the analytical column A fourth flow path for allowing the suction / discharge means to suck the cleaning liquid or the regeneration liquid, and a fourth flow for supplying the cleaning liquid or the regeneration liquid discharged from the suction / discharge means to the first flow path switching means. a second flow path switching unit for switching between road, and controls the operation of the liquid feeding means, the sample introduction means, a solution suction discharge means, the first flow path switching unit and the second flow path switching unit, the first During the period when the flow path of the flow path switching unit is in the first flow path state, the flow path of the second flow path switching unit is set to the third flow path state so that the cleaning liquid or the regeneration liquid is the solution suction and discharge means. And after the flow path of the first flow path switching unit is set to the fourth flow path state during the period in which the flow path of the first flow path switching means is in the second flow path state, Alternatively, the regeneration solution is introduced into the sample introduction unit, and the cleaning solution or the regeneration solution is introduced into the sample introduction unit. The channel of one channel switching unit is switched to the first channel state, the channel of the second channel switching unit is switched to the third channel state, and introduced into the sample introduction unit. And a control means for supplying the washing liquid or the regenerating liquid to the analytical column .

(2) Preferably, in (1) above, the control means is configured such that the flow path of the second flow path switching means is in a period during which the flow path of the first flow path switching means is in the first flow path state. Is introduced into the solution suction and discharge means as the third flow path state, and the second flow is changed during the period in which the flow path of the first flow path switching means is in the second flow path state. With the flow path of the path switching means as the fourth flow path state, the second flow path switching means is switched to the third flow path state with the cleaning liquid or the regeneration liquid introduced into the sample introduction means, After the cleaning liquid or the regeneration liquid is sucked by the solution suction / discharge means, the flow path of the second flow path switching means is set to the fourth flow path state, and the cleaning liquid or the regeneration liquid is further introduced into the sample introduction means. The cleaning liquid or the regenerating liquid is introduced into the sample introduction means, and the cleaning liquid or the regenerating liquid is the sample. The flow path of the first flow path switching means is switched to the first flow path state and the flow path of the second flow path switching means is changed to the third flow path state in a state where the flow path is introduced into the input means. By switching, the cleaning liquid or the regenerating liquid introduced into the sample introduction means is supplied to the analytical column.

(3) Preferably, in the above (1), the control means sets the flow path of the second flow path switching means during the period when the flow path of the first flow path switching means is in the first flow path state. As the third flow path state, a cleaning liquid or a regenerative liquid is introduced into the solution suction / discharge means, and the second flow path is in a period during which the flow path of the first flow path switching means is in the second flow path state. With the flow path of the switching means as the fourth flow path state, the cleaning liquid or the regeneration liquid is introduced into the sample introduction means, and the first flow path switching is performed with the cleaning liquid or the regeneration liquid introduced into the sample introduction means. The flow path of the means is switched to the first flow path state, the flow path of the second flow path switching means is switched to the third flow path state, and the cleaning liquid or regenerative liquid introduced into the sample introduction means Is supplied to the analytical column, and the flow path of the first flow path switching means is switched to the second flow path state. A state in which the flow path of the second flow path switching unit is set to the fourth flow path state, the cleaning liquid or the regeneration liquid is further introduced into the sample introduction means, and the cleaning liquid or the regeneration liquid is further introduced into the sample introduction means The sample introduction unit is configured to switch the channel of the first channel switching unit to the first channel state, switch the channel of the second channel switching unit to the third channel state, and Is supplied to the analytical column.
(4) In the liquid chromatographic analysis method, a fixed amount of the solution can be held, and the solution from the liquid feeding means for feeding the solution is introduced into the sample introduction means for introducing the sample to be measured into the solution. And supplying the measurement target sample together with the solution from the sample introduction means to the analysis column, and sucking the cleaning liquid or the regenerating liquid from the sample introduction means during the period during which the measurement target sample and the solution are supplied to the analysis column, The solution suction / discharge means that can hold and discharge a certain amount of the liquid sucks and holds a fixed amount of the cleaning liquid or the regenerated liquid, and directly supplies the solution from the liquid feeding means to the analysis column without passing through the sample introduction means. the period for supplying said cleaning liquid or regeneration liquid held in the solution suction discharge unit, is held is supplied to the sample introduction means, is held in the sample introduction means And the cleaning liquid or regenerated liquid is supplied to the analytical column to wash or regeneration of the analytical column.
(5) Preferably, in (4) above, after the solution from the liquid feeding means is directly supplied to the analytical column without going through the sample introduction means, the washing liquid or the regenerating liquid held in the solution suction / discharge means Is supplied to and held in the sample introduction means, and the solution suction and discharge means is further sucked and held by a fixed amount, and the cleaning or regeneration liquid held in the solution suction and discharge means is The sample introduction means is supplied and held, and the cleaning liquid or the regeneration liquid held in the sample introduction means is supplied to the analysis column, and the analysis column is washed or regenerated.
(6) Preferably, in the above (4), the cleaning liquid or the regenerating liquid held in the sample introduction means is supplied to the analytical column to clean or regenerate the analytical column, and the solution suction / discharge The cleaning liquid or the regenerating liquid is further sucked and held in the means, and the cleaning liquid or the regenerating liquid held in the solution suction / discharge means is supplied to and held in the sample introducing means, and is held in the sample introducing means. The washed or regenerated solution is supplied to the analytical column, and the analytical column is further washed or regenerated.

  According to the present invention, it is possible to realize a liquid chromatographic analysis method and apparatus capable of regenerating or washing an analytical column in a short time even with an isocratic liquid feeding method, while having an inexpensive configuration.

  That is, according to the present invention, by selecting an arbitrary column regeneration solution or washing solution and introducing the temporarily retained column regeneration solution or washing solution into the analysis flow path, without a gradient system, The analytical column can be automatically regenerated or washed in a short time.

Hereinafter, an embodiment of the present invention will be described with reference to the accompanying drawings.
FIG. 1 is a flow chart for introducing a regenerating solution of an analytical column into a syringe during a period of introducing the sample into the analytical column in the liquid chromatograph analyzer according to one embodiment of the present invention.

  In FIG. 1, a pump (not shown) and an analysis column (not shown) are connected to an injection valve 11. The injection valve 11 has ports 111 to 116, and the flow path can be switched. The port 111 of the injection valve 11 is connected to the pump, and the port 112 is connected to the analytical column.

  The port 116 of the injection valve 11 is connected to the port 113 through the sample loop 10, the needle 17, and the injection port 16.

  Moreover, the syringe valve 12 has ports S1 to S6, and the flow path can be switched. The port S2 of the syringe valve 12 is connected to the port 114 of the injection valve 11, and the port S3 of the syringe valve 12 is connected to the port 115 of the injection valve 11.

  The port S4 of the syringe valve 12 is connected to the cleaning port 15, and the port S5 is connected to a flow path for pump cleaning. The port S6 of the syringe valve 12 is connected to the syringe 14, and the port S1 is connected to the solvent switching valve 13.

  The solvent switching valve 13 can supply any one of the cleaning liquid 18, the regeneration liquid 19, and the needle cleaning liquid 20 to the syringe valve 13.

  Although not shown, the liquid chromatograph apparatus includes control means for controlling operations of the injection valve 11, the syringe valve 12, the syringe 14, the solvent switching valve 13, a pump, and the like.

  In the above configuration, the eluent from the pump (not shown) is analyzed through the port 111 and port 116 of the injection valve 11, the sample loop 10, the needle 17, the injection port 16, the port 113 and port 112 of the injection valve 11. It is introduced into a column (not shown).

  During the period when the eluent from the pump is introduced into the analytical column, the switching valve 13 is switched to the regenerating liquid 19 side, and the flow path to which the ports S1 and S6 of the syringe valve 12 are connected is selected. . Then, as described above, the syringe 14 is aspirated during the period in which the eluent from the pump is introduced into the analysis column, and the regenerating liquid 19 is filled into the syringe 14 via the solvent switching valve 13.

  FIG. 2 shows that after the regeneration solution 19 is filled in the syringe 14, the regeneration solution is introduced from the syringe 14 into the sample loop during a period in which the solution from the pump is supplied to the analytical column without passing through the sample loop 10. FIG.

  After the ports S6 and S1 connected to the syringe valve 12 are switched to the ports S6 and S2, the position of the injection valve 11 is switched to the load side.

  Thereby, the solution from the pump is supplied to the analytical column via the ports 111 and 112. Also, port S6 and port S2 of the syringe valve 12 connected to the syringe 14, port 114 and port 113 of the injection valve 11, injection port 16, needle 17, sample loop 10, port 116, port 115 and port of the syringe valve 12 A flow path passing through S3, port S4, and washing port 15 is formed. In this state, the inside of the sample loop 10 is an eluent.

  In the above flow path, the regeneration liquid 19 in the syringe 14 can be pushed out and the inside of the sample loop 10 can be replaced with the regeneration liquid 19.

  FIG. 3 is a flow chart for introducing the column regeneration solution in the loop into the analysis column after replacing the inside of the sample loop 10 with the regeneration solution 19. Basically, it is the same flow path as the flow path shown in FIG.

  After switching so that the ports S2 and S3 of the syringe valve 12 communicate with each other, the position of the injection valve 11 is switched to the Inject side.

  As a result, the regeneration solution 19 in the loop is introduced into the column through the port 116 of the injection valve 11, the sample loop 10, the needle 17, the injection port 16, the port 113, and the port 112.

  FIG. 4 is a flow chart when the column regenerating solution introduced into the sample loop in the state shown in FIG. 2 alone is not sufficient, and the column regenerating solution is additionally introduced into the syringe 14. is there. Since the amount of the regenerating liquid in the analytical column changes depending on the size of the analytical column, a large amount of the regenerating liquid can be obtained by increasing the capacity of the sample loop 10 and introducing the regenerating liquid 19 from the syringe 14 a plurality of times. 19 can be introduced into the analysis channel.

In addition, the syringe 14 is in a suction state through the flow path shown in FIG.

  Next, the flow path shown in FIG. 3 is switched to the flow path shown in FIG. 2, and after switching so that the ports S6 and S2 of the syringe valve 12 communicate with each other, the position of the injection valve 11 is switched to the load side.

  As a result, the flow path passes through the syringe 14, the port S6, the port S2, the port 114, the port 113, the injection port 16, the needle 17, the sample loop 10, 116, the port 115, the port S3, the port S4, and the washing port 15.

  In this state, since the inside of the sample loop 10 is an eluent, the regenerating liquid 19 in the syringe 14 is pushed out, and the inside of the sample loop 10 is replaced with the regenerating liquid 19.

  Thereafter, the analysis column is regenerated using the flow path shown in FIG.

  In the above description, the regeneration liquid is supplied from the solvent switching valve 13 to the syringe valve 12. However, the cleaning liquid 18 and the needle cleaning liquid 20 are supplied to the syringe valve 12 by switching the solvent switching valve 13. Is possible.

  The cleaning port 15 is a port for cleaning the outer wall of the needle 17.

FIG. 5 is a schematic configuration diagram of a liquid chromatograph analyzer according to an embodiment of the present invention.
In FIG. 5, a liquid chromatograph apparatus includes a pump 22 for feeding an eluent 21, an autosampler 23 for introducing a sample into a flow path, an analysis column 30 for separating components, and the analysis column. A column oven 24 for constant temperature 30 and a detector 25 for detecting the eluted components from the column 30 are provided.

  Further, a column cleaning liquid 27, a regenerating liquid 28, a needle cleaning liquid 29 for cleaning the sample, and the like are supplied to the analysis column 30 by the autosampler 23.

  The configuration shown in FIGS. 1 to 4 corresponds to the autosampler 23 having the configuration shown in FIG. The autosampler 23 includes control means for controlling the operations of the injection valve 11, the syringe valve 12, the syringe 14, and the solvent switching valve 13 shown in FIG. Further, the waste liquid 26 is discharged from the detector 25.

  FIG. 6 is a graph comparing a chromatogram obtained by a general liquid chromatograph analyzer with a chromatogram obtained by a liquid chromatograph device according to the present invention. The vertical axis of the graph shown in FIG. 6 indicates the signal intensity, and the horizontal axis indicates the holding time. 6A is a chromatogram obtained by a conventional liquid chromatograph, and FIGS. 6B and 6C are chromatograms obtained by a liquid chromatograph according to the present invention. is there. The chromatogram shown in (C) of FIG. 6 is the result obtained when the column regenerating solution was introduced earlier than the column regenerating solution introduction timing from which the chromatogram shown in (B) of FIG. 6 was obtained. It is.

  In the case of the isocratic elution method, impurities may be eluted even after the target peak component is eluted. Each of (A), (B), and (C) in FIG. 6 indicates this. The end of the analysis is a time when it can be determined that the detection of the impurity has been completed.

  In the chromatogram shown in FIG. 6A, the end point of analysis is t3, t2 in FIG. 6B, and t1 in FIG. 6C, and the analysis of the liquid chromatograph apparatus according to the present invention is performed. The end points t1 and t2 are shorter than the analysis end point t3 of the liquid chromatograph apparatus according to the prior art, and the analysis time is shortened.

  This is because in the present invention, after the elution of the target peak is confirmed, the analysis cycle is accelerated by introducing a regenerating solution into the column. That is, the sample introduction, washing, and column regeneration are performed in the analysis column without stopping the operation during one analysis, and thus the analysis cycle can be shortened.

  Here, the chromatogram shown in FIG. 6C is a simulated chromatogram when the timing for introducing the column regenerating solution is set earlier as described above. By calculating the dead volume from the injection valve 11 to the detector 25, the analysis cycle can be further shortened by introducing the regenerated solution of the column into the analysis flow path without confirming the elution of the target peak. This is an example where this is possible.

  As described above, according to the present invention, the autosampler device is used, and during the sample introduction period into the analytical column, the syringe 14 is filled with the regenerating liquid or the cleaning liquid, and components other than the target component of the analytical column are removed. During the process and the process of introducing the target component into the detector, the sample loop is filled with the cleaning liquid filled in the syringe 14. Then, in the next analytical column regeneration step, the regeneration solution filled in the sample loop is introduced into the analysis column.

  Therefore, it is possible to realize a liquid chromatographic analysis method and apparatus capable of regenerating or washing an analysis column in a short time while using an inexpensive configuration without using an expensive gradient system.

FIG. 3 is a flow chart for introducing the regenerated solution of the analytical column into the syringe during the period of introducing the sample into the analytical column in the liquid chromatograph analyzer according to one embodiment of the present invention. FIG. 4 is a flow chart for introducing the regenerative liquid from the syringe into the sample loop after the regenerative liquid is filled in the syringe in the liquid chromatograph analyzer according to the embodiment of the present invention. FIG. 3 is a flow chart for introducing the column regenerating solution in the loop into the analytical column after replacing the sample loop with the regenerating solution in the liquid chromatograph analyzer according to one embodiment of the present invention. FIG. 3 is a flow chart when a column regenerating solution is additionally introduced into a syringe in the liquid chromatograph analyzer according to an embodiment of the present invention. It is a schematic block diagram of the liquid chromatograph analyzer which is one Embodiment of this invention. It is a graph which compares the chromatogram obtained by the general liquid chromatograph apparatus with the chromatogram obtained by the liquid chromatograph apparatus by this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 10 Sample loop 11 Injection valve 12 Syringe valve 13 Solvent switching valve 14 Syringe 15 Washing port 16 Injection port 17 Needle 18, 27 Washing liquid 19, 28 Regeneration liquid 20, 29 Needle washing liquid 21 Elution liquid 22 Pump 23 Autosampler 24 Column oven 25 Detection 26 Waste liquid 30 Analytical column

Claims (6)

In a liquid chromatograph analyzer,
A liquid feeding means for feeding the solution;
A sample introduction means capable of holding a certain amount of the solution and introducing the sample to be measured into the solution;
An analytical column that adsorbs and elutes the sample to be measured;
Detection means for detecting the sample to be measured eluted from the analytical column;
A solution suction / discharge means capable of sucking a cleaning liquid or a regenerating liquid, holding a fixed amount, and discharging it;
A first flow path state in which the solution fed from the liquid feeding means is supplied to the analytical column via the sample introduction means, and the solution sent from the liquid feeding means is passed through the sample introduction means. First flow path switching means for switching between a second flow path state for supplying the cleaning liquid or the regenerating liquid discharged from the suction / discharge means to the sample introduction means without being supplied to the analysis column,
A third flow path for sucking the cleaning liquid or the regeneration liquid to the suction / discharge means; and a fourth flow path for supplying the cleaning liquid or the regeneration liquid discharged from the suction / discharge means to the first flow path switching means. A second flow path switching means for switching;
The operation of the liquid feeding means, sample introduction means, solution suction / discharge means, first flow path switching means and second flow path switching means is controlled, and the flow path of the first flow path switching means is the first flow path switching means. During the period of the flow path state, the flow path of the second flow path switching means is set to the third flow path state to introduce the cleaning liquid or the regeneration liquid into the solution suction / discharge means, and the flow path of the first flow path switching means In the period of the second flow path state, the flow path of the second flow path switching unit is set to the fourth flow path state, and the cleaning liquid or the regenerating liquid is introduced to the sample introduction unit. The channel of the channel switching unit is switched to the first channel state, the channel of the second channel switching unit is switched to the third channel state, and introduced into the sample introduction unit. Control means for supplying a washing solution or a regenerating solution to the analytical column;
A liquid chromatograph analyzer comprising:   2. The liquid chromatograph analyzing apparatus according to claim 1, wherein the control means is configured to cause the flow of the second flow path switching means during a period in which the flow path of the first flow path switching means is in the first flow path state. A cleaning liquid or a regenerating liquid is introduced into the solution suction / discharge means with the path set as the third flow path state, and the second flow path state is maintained while the flow path of the first flow path switching means is in the second flow path state. With the flow path of the flow path switching means as the fourth flow path state, the second flow path switching means is switched to the third flow path state with the cleaning liquid or the regeneration liquid introduced into the sample introduction means. After the cleaning liquid or the regenerating liquid is sucked by the solution suction / discharge means, the flow path of the second flow path switching means is set to the fourth flow path state, and the cleaning liquid or the regenerating liquid is further introduced into the sample introducing means. And the cleaning liquid or the regenerating liquid is introduced into the sample introduction means. The channel of one channel switching unit is switched to the first channel state, the channel of the second channel switching unit is switched to the third channel state, and introduced into the sample introduction unit. A liquid chromatograph analyzer, wherein the cleaning liquid or the regenerated liquid is supplied to the analytical column.   2. The liquid chromatograph analyzing apparatus according to claim 1, wherein the control means is configured to cause the flow of the second flow path switching means during a period in which the flow path of the first flow path switching means is in the first flow path state. A cleaning liquid or a regenerating liquid is introduced into the solution suction / discharge means with the path set as the third flow path state, and the second flow path state is maintained while the flow path of the first flow path switching means is in the second flow path state. With the flow path of the flow path switching means as the fourth flow path state, the cleaning liquid or the regeneration liquid is introduced into the sample introduction means, and the first flow is introduced in the state where the cleaning liquid or the regeneration liquid is introduced into the sample introduction means. The flow path of the path switching means is switched to the first flow path state, the flow path of the second flow path switching means is switched to the third flow path state, and the cleaning liquid introduced into the sample introduction means or After supplying the regenerating solution to the analytical column, the flow path of the first flow path switching means is moved to the second flow path. After switching to the state and setting the flow path of the second flow path switching means to the fourth flow path state, the cleaning liquid or the regeneration liquid is further introduced into the sample introduction means, and the cleaning liquid or the regeneration liquid is introduced into the sample introduction means. Further, the flow path of the first flow path switching means is switched to the first flow path state in the introduced state, and the flow path of the second flow path switching means is switched to the third flow path state. A liquid chromatograph analyzing apparatus, wherein the cleaning liquid or the regenerating liquid introduced into the sample introduction means is supplied to the analytical column. In the liquid chromatographic analysis method,
A constant amount of solution can be held, and the solution from the liquid feeding means for feeding the solution is introduced into the sample introduction means for introducing the measurement target sample into the solution.
Supplying the sample to be measured together with the solution from the sample introduction means to the analytical column;
During the period in which the sample to be measured and the solution are supplied from the sample introduction means to the analytical column, the cleaning liquid or the regenerating liquid is sucked into the solution aspirating and discharging means that can suck and hold a certain amount of the cleaning liquid or the regenerating liquid. Suck a certain amount and hold it,
During the period in which the solution from the liquid feeding means is directly supplied to the analytical column without going through the sample introduction means, the cleaning liquid or the regenerating liquid held in the solution suction / discharge means is supplied to and held in the sample introduction means. Let
A liquid chromatographic analysis method, wherein a cleaning liquid or a regenerating liquid held in the sample introduction means is supplied to the analytical column, and the analytical column is cleaned or regenerated. The liquid chromatograph analysis method according to claim 4,
After supplying the solution from the liquid feeding means directly to the analytical column without going through the sample introduction means,
The cleaning liquid or the regeneration liquid held in the solution suction / discharge means is supplied to and held in the sample introduction means,
The solution suction / discharge means holds the cleaning liquid or the regenerating liquid by sucking a certain amount further,
The cleaning liquid or the regeneration liquid held in the solution suction / discharge means is further supplied to the sample introduction means to be held,
A liquid chromatographic analysis method, wherein a cleaning liquid or a regenerating liquid held in the sample introduction means is supplied to the analytical column, and the analytical column is cleaned or regenerated. The liquid chromatograph analysis method according to claim 4,
The cleaning liquid or the regenerating liquid held in the sample introduction means is supplied to the analysis column, and the analytical column is cleaned or regenerated, and the solution suction and discharge means is further sucked with a certain amount of the cleaning liquid or the regenerating liquid. Hold
The cleaning liquid or the regeneration liquid held in the solution suction / discharge means is further supplied to the sample introduction means to be held,
A liquid chromatographic analysis method characterized in that a cleaning liquid or a regenerating liquid held in the sample introduction means is supplied to the analytical column, and the analytical column is further cleaned or regenerated.

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