JP2839687B2 - Electrophoresis-mass spectrometer - Google Patents

Description

The present invention relates to an analyzer configured by connecting an electrophoresis apparatus and a mass spectrometer online.

[Prior art] Recently, an electrophoresis apparatus and a mass spectrometer are connected online, a sample solution separated by the electrophoresis apparatus is introduced into an ionization chamber of the mass spectrometer, and a neutral particle beam collides with the sample to ionize the sample. An example of how to do this is "RAPID COMMUNICATIO
NS IN MASS SPECTROMETRY.VOL 3.NO 10.1989. (348p-3
51p)].

The device introduced in this document has a configuration as shown in FIG.

In FIG. 4, reference numeral 1 denotes a mobile phase storage container storing a mobile phase for electrophoresis, 2 denotes a high-voltage power supply for applying a positive high voltage to the mobile phase in the mobile phase storage container 1, and 3 denotes an electrophoresis mobile phase. One end of the separation column is inserted into the mobile phase. The sample liquid separated by the separation column 3 is introduced into an ion source (FAB ion source) via an introduction tube 4 composed of a capillary tube or the like, and is ionized by neutral particle beam impact. The other end of the separation column 3 is inserted into the matrix through a wall of a matrix storage container 6 that stores a matrix such as glycerin that promotes ionization of a sample. Then, the introduction pipe 4 is inserted into the storage container 6 from the opposite direction, and the tip of the introduction pipe 4 and the tip of the separation column 3 are arranged so as to face each other at an appropriate interval. The matrix in the matrix container 6 is electrically grounded.

In such a configuration, one end inserted into the mobile phase of the separation column 3 is transferred to the sample solution and lifted several cm to several tens of seconds higher than the other end of the separation column by several cm.
A sample is introduced into a separation column by a method of introducing a sample using a head, or a method of introducing an electrode into a sample solution by electrophoresis and electroosmosis. Thereafter, one end of the separation column 3 is returned into the mobile phase to which the positive high voltage has been applied. Then, the introduced sample is separated under the effects of both electroosmosis and electrophoresis, and is separated into the separation column 3 inserted in the matrix solution stored in the matrix storage container 6 and electrically grounded. Move towards the other end of the. An opening at one end of the introduction tube 4 is opposed to the opening at the other end of the separation column 3 at an appropriate interval, for example, about several tens of μm. Since the opening at the other end of the introduction tube 4 is set in the ion source 5 in a vacuum atmosphere, the separated sample that has reached the opening at the other end of the separation column 3 is introduced together with the matrix at atmospheric pressure. It is sucked into the ion source 5 through the tube 4. The sample and the matrix introduced into the ion source 5 in this manner are ionized, introduced into a mass spectrometer (not shown), and subjected to mass analysis.

[Problems to be Solved by the Invention] However, in order to measure a different sample, even if there is a demand to quickly change the type of matrix held in the matrix storage container and perform mass spectrometry, the ion source is required. The absolute flow rate of the matrix that can be introduced is very small, and it takes a very long time to remove the previously used matrix, and the matrix cannot be completely removed. Was difficult.

The present invention has been made in view of the above points, and has as its main object to provide an apparatus capable of satisfying a demand for mass spectrometry by rapidly changing the type of matrix in order to measure a different sample. Purpose.

[Means for Solving the Problems] In order to achieve the above object, the present invention provides an electrophoresis apparatus having a separation column, a mass spectrometer, and a mass spectrometer which uses a sample solution separated by the separation column of the electrophoresis apparatus. And a mixing section provided in the middle of the introduction path for adding the additive liquid to the sample solution. The mobile phase supplied to the column and the additive liquid supplied to the mixing section have the same variable pressure. And a discharge channel for discharging the additive liquid from the mixing section is provided.

Embodiment An embodiment of the present invention will be described below in detail with reference to the accompanying drawings.

In FIG. 1, the same components as those of the conventional apparatus shown in FIG. 4 are denoted by the same reference numerals. The device of FIG.
The different point of the apparatus shown in the figure is that an opening 15 is provided at the bottom of the matrix container 6 and a discharge passage having an on-off valve 16 in the middle.
17 is provided.

In such a configuration, first, the on-off valve 16 provided at the bottom of the matrix storage container 6 is closed, and then the matrix is injected. After the sample is introduced by the method described in the conventional example, the sample that has been separated into three by the separation column by the method described in the proposed apparatus is sucked into the ion source 5 through the introduction tube 13 together with the matrix. The sample and the matrix introduced into the ion source 5 in this manner are ionized, introduced into a mass spectrometer (not shown), and subjected to mass analysis.

Next, when the type of matrix is changed in order to measure a different sample, when the on-off valve 16 is opened, the matrix held in the matrix storage container 6 is discharged from the discharge channel 17. After the matrix has been completely discharged, the on-off valve 16 is closed, and then another type of matrix is injected. Then, the sample is measured as described above.

FIG. 2 shows the electrophoresis described in Japanese Patent Application No. 2-226064.
An embodiment apparatus in which the present invention is applied to a mass spectrometer is shown. 2, the same components as those of the conventional example shown in FIG. 4 are denoted by the same reference numerals. In FIG. 2, reference numeral 7 denotes a sealed mobile phase storage container having an opening in which a mobile phase for electrophoresis is held in advance and gas pressure can be applied to the mobile phase. The mobile phase stored in the mobile phase storage container 7 is a high-voltage power supply 2.
, A positive high voltage is applied. Numeral 8 denotes a sealed matrix container connected to a matrix reservoir 9 having a matrix pre-stored therein and having an opening through which gas pressure can be applied to the matrix. The matrix stored in the matrix storage container 8 is electrically grounded. 10 is a gas cylinder for supplying gas pressure applied to the mobile phase and the matrix, 11 is a pressure regulating valve, and 12 is a gas cylinder 10
These are Teflon pipes connected to the respective openings for introducing the gas supplied from the above into the mobile phase storage container 7 and the matrix reservoir 9. Numeral 13 denotes an inlet tube which is appropriately thinner than the inner diameter of the inlet tube described in the conventional example in order to increase the flow resistance. The introduction tube 13 has one end inserted into the matrix container 8 and the other end inserted into the FAB ion source 5 as described in the conventional example. Then, the separation column 3 and the introduction tube 13 are inserted from both sides of the matrix storage container 8, and as described in the conventional example, the open end of the separation column 3, the introduction tube 13 and the open end face each other with an appropriate distance therebetween. Are located. A guide 14 is provided near the opening end of the introduction tube 13 so that the center of the opening of the separation column 3 does not deviate from the center of the opening of the introduction tube 13. Further, the separation column 3 and the Teflon pipe 12 are electrically insulated.

In such a configuration, after introducing the sample by the method described in the conventional example, one end of the separation column 3 is returned to the mobile phase to which a positive high voltage is applied. Then, the gas cylinder is opened, and the gas pressure adjusted to an appropriate pressure is applied to the mobile phase in the mobile phase storage container 7 and the matrix in the matrix reservoir 9.
Therefore, the introduced sample moves toward the other end of the separation column 3 inserted into the matrix solution stored in the matrix storage container 8 while being separated by the effects of both electroosmosis and electrophoresis. . And this introduction pipe
Since the opening at the other end of the column 13 is set in the ion source 5 in a vacuum atmosphere, the separated sample that has reached the opening at the other end of the separation column 3 is removed together with the matrix to which an appropriate gas pressure is applied. It is sucked into the ion source 5 through the introduction tube 13. The sample and the matrix introduced into the ion source 5 in this manner are ionized, introduced into a mass spectrometer (not shown), and subjected to mass analysis.

Next, when changing the type of matrix to measure a different sample, open the on-off valve 16 and then open the gas cylinder.
From 10 a gas pressure is applied to the matrix reservoir 9. The matrix held in the matrix reservoir 9 and the matrix storage container 8 is discharged from the discharge channel 17. After all the matrix has been discharged, the pressure regulating valve 11 is closed to stop applying the gas pressure. Then, the on-off valve 16 is closed, and then the Teflon pipe connected to the opening of the matrix reservoir 9
Remove 12 and inject another type of matrix through the opening. Thereafter, the Teflon pipe 12 is attached to the opening of the matrix reservoir 9 again. Then, the sample is measured as described above.

In the present embodiment, the Teflon pipe 12 connected to the opening of the matrix reservoir 9 is removed and the matrix is injected from the opening. However, the present invention is not limited to this. A valve into which the matrix can be injected, or an injection port closed with a rubber stopper or the like so that the matrix in the syringe can be injected through the injection needle may be provided.

FIG. 3 is a schematic diagram showing the configuration of an apparatus according to another embodiment of the present invention. 3, the same components as those of the embodiment shown in FIG. 2 are denoted by the same reference numerals. The difference between the apparatus of FIG. 3 and the apparatus of FIG. 2 is that matrix storage containers 18 and 19 holding different types of matrices,
Pumps 20 and 21 for transferring the matrix held in the matrix storage containers 18 and 19 to the matrix reservoir 9 and a switching valve 22 for switching the flow path are provided.

In such a configuration, first, the on-off valve 16 is closed in a state where no matrix remains in the matrix storage container 8 and the matrix reservoir 9, and then, in a state where gas pressure is not applied to the matrix in the matrix reservoir 9. ,
After switching the switching valve 22 to A, the pump 20 is operated to transfer the matrix held in the matrix container 18 to the matrix container 8 and the matrix reservoir 9. Thereafter, the sample is measured as described above.

Next, when changing the type of matrix to measure a different sample, open the on-off valve 16 and then open the gas cylinder.
From 10 a gas pressure is applied to the matrix reservoir 9. After discharging the matrix as described above,
Close 11 to stop applying gas pressure. Then, the on-off valve 16 is closed. Then, after switching the switching valve 22 to B, the pump 21
Is operated to transfer the matrix held in the matrix storage container 19 to the matrix storage container 8 and the matrix reservoir 9. Then, as described above,
Measure the sample.

In the present embodiment, two types of storage containers holding different types of matrices are prepared so that different matrices can be selected. However, the present invention is not limited to this, and three or more types of different matrices are prepared. Alternatively, it may be possible to select with a switching valve.

At this time, the opening and closing of the discharge valve may be switched by an electromagnetic valve, and the switching valve may be switched by a motor or the like, so that the matrix can be replaced automatically.

Further, in the proposed device, the mobile phase storage container and the matrix storage container are pressurized together using one pressure regulating valve. You may pressurize separately.

In addition, in the proposed device, a matrix reservoir connected to the matrix storage container is provided and gas pressure is applied.However, without providing a matrix reservoir, a sufficient amount of matrix for measurement can be held, and gas pressure is applied to the matrix. A matrix container having an opening that can be provided may be provided.

Further, in the proposed device, the matrix is added to the sample solution in the mixing section and introduced into the FAB ion source, and ionized.However, the present invention is not limited to this.
For example, water, methanol, acetone, acetonitrile, or the like may be added to the sample solution from the mixing section, and introduced into a chemical ionization (CI) ion source to ionize the sample.
In this case, the pressure in the ionization chamber is maintained at, for example, about 1 Torr suitable for chemical ionization, and the sample component is chemically ionized using the additive liquid or the mobile phase gas vaporized in the ionization chamber as a reaction gas. At this time, if the pressure in the ionization chamber is set sufficiently low, ionization by electron impact (EI) can be performed.

[Effects] As described in detail above, according to the present invention, the sample solution separated by the separation column of the electrophoresis apparatus is added to the sample liquid in the mixing section provided in the introduction path for introducing the sample solution into the mass spectrometer. By providing a means for introducing the additive liquid supplied for and a discharge flow path for discharging the additive liquid from the mixing section,
A device that can easily remove the additive solution and introduce another additive solution, so that it can satisfy the need to quickly change the type of matrix and perform mass spectrometry in order to measure different samples. Can be provided.

[Brief description of the drawings]

FIG. 1 is a schematic diagram showing the configuration of an embodiment of the present invention, and FIG.
FIG. 3 and FIG. 3 are schematic diagrams showing the configuration of an apparatus according to another embodiment of the present invention, and FIG. 4 is a schematic view showing the configuration of a conventional apparatus. 1,7: Mobile phase storage container 2: High pressure power supply, 3: Separation column 4, 13: Introductory tube, 5: FAB ion source 6, 8, 18, 19: Matrix storage container 9: Matrix reservoir 10: Gas cylinder, 11: Pressure control valve 12: Teflon pipe, 14: Guide 15: Opening, 16: On / off valve 17: Drain flow path 20, 21: Pump, 22: Switching valve

──────────────────────────────────────────────────続 き Continued on the front page (58) Field surveyed (Int.Cl. ⁶ , DB name) G01N 27/62-27/70 G01N 27/447-27/453 H01J 49/00-49/48

Claims (2)

(57) [Claims] 1. An electrophoresis apparatus having a separation column, a mass spectrometer, an introduction path for introducing a sample solution separated by the separation column of the electrophoresis apparatus into the mass spectrometer, and an additive liquid for adding the additive solution to the sample solution. A mixing section provided in the middle of the introduction path, wherein the same variable pressure gas pressure is applied to the mobile phase supplied to the column and the additive liquid supplied to the mixing section, and added from the mixing section. An electrophoresis-mass spectrometer comprising a discharge channel for discharging a liquid. 2. An additive liquid reservoir for storing an additive liquid to be supplied to the mixing section, and an additive liquid switching supply means for selectively supplying different types of additive liquid to the additive liquid reservoir. The electrophoresis-mass spectrometer according to claim 1, wherein: