JPS60138450A - Thin tube type isotachophoresis analysis - Google Patents

Abstract

PURPOSE:To prevent the formation of a mixed zone of both liquids by monitoring the introduction of a terminal liquid and a leading liquid introduced thereafter at an exhausting hole provided deviatedly to an electrophoresis tube. CONSTITUTION:The terminal electrolyte T is introduced from a terminal liquid electrode cell 2. Since the exhausting hole 7 provided deviatedly to the electrophoresis tube 3 is opened, the electrolyte T is exhausted from the hole 7 in order, and the terminal electrolyte introduction is stopped when the exhausting is attained. Next, the leading electrolyte L is introduced from a leading liquid electrode cell 4, said electrolyte is attained to the hole 7 and exhausted partially together with the terminal electrolyte. At the exhausting time point, the hole 7 is closed by an on-off valve 9. Thereby, a boundary surface P of both electrolytes is formed clearly. The sample is injected from a sample injecting hole 6 by using a microsyringe 11. Since a needle 12 of the microsyringe 11 is not passed through the terminal electrolyte at all, the mixed zone of both electrolytes is not formed.

Description

[Detailed description of the invention] This invention relates to a capillary isotachophoresis analysis method.

More specifically, this invention introduces a terminal electrolyte and a leading electrolyte into the migration I tube from the terminal liquid electrode tank side and the leading liquid electrode tank side, respectively, and drains these two electrolytes with a pressurized on-off valve. A capillary type, etc. in which the introduction of both electrolytes is stopped when the electrolytes are discharged from the mouth, and then the sample injected into the electrophoresis capillary from the sample injection port is subjected to isokinetic electrophoresis to detect the target substance in the sample. In fast electrophoresis analysis, the terminal electrolyte is introduced from the terminal liquid electrode tank into the electrophoresis capillary, and the electrolyte includes the position of the sample injection port, and the drain port is located unevenly in the migration tube on the leading liquid electrode tank side. Stop the introduction as soon as it is ejected from the system, and after that stop, the leading! This invention relates to a capillary isokinetic electrophoresis analysis method in which a terminal electrolyte is introduced from an electrode bath into an electrophoresis capillary.

In the electrophoresis method, a communicating electrophoresis tube (key 17 cell tube) is filled with a terminal electrolyte solution and a gain gummy solution, and substances (amino acids, peptides, biological substances, etc.) that become charged at the interface are charged. There is a capillary isokinetic electrophoresis method in which a sample is injected, electrophoresis is performed using a constant current, and the substance to be detected is separated (fractionation) for qualitative and/or quantitative determination. In this electrophoresis method, a typical sample injection part is
For example, as shown in Fig. 3, the bent part (5a) is configured so that the needle (12a) of the microsyringe (11a) extends toward the leading liquid electrode tank (4a) when injecting the sample, and the drain port is also connected to the sample inlet. Leading liquid electrode tank (4a
) are placed apart from each other. Therefore, an interface between both electrolytes is formed near the drain port, and when injecting a sample, the needle of the microsyringe passes through the terminal electrolyte and reaches the interface or leading electrolyte, and when the needle is inserted and removed, both Easy to create a mixing zone between electrolyte and sample. As a result, the electrophoresis distance required to separate both electrolytes is required at the stage prior to sample separation, and a considerably long electrophoresis tube and electrophoresis time are required.

This invention was developed in view of these circumstances, and its main features are that the terminal electrolyte is introduced into the electrophoresis capillary from the terminal liquid electrode tank, and that the electrolyte includes the position of the sample injection port and the leading liquid electrode tank. This feature lies in the fact that the introduction of the electrolyte was stopped when the liquid was discharged from the drain port located unevenly into the electrophoresis tube on the layer side, and after that, the terminal electrolyte was introduced from the leading liquid electrode tank into the electrophoresis tube. Therefore, when injecting a sample, a nail such as a microsyringe can pass only through the leading electrolyte, so there is little mixing with the terminal electrolyte, and the migration distance and time can be shortened. As a specific example of a structure in which the drain port of the electrophoresis tube is located at the position of the sample injection port and is located unevenly in the electrophoresis tube on the leading liquid electrode tank side, the example of the embodiment is mentioned as a preferable example, but there are other examples. One example is one in which a linear electrophoresis capillary section that is not bent is provided facing the sample injection port. In each of these examples, the terminal electrolyte is first introduced, and then the leading electrolyte is introduced.

The present invention will be described in detail below based on embodiments shown in the figures. Note that this invention is not limited to this.

First, in FIG. 1, a capillary electrophoresis analyzer (1) showing an example of a device for carrying out the present invention consists of a terminal liquid electrode tank (21), an electrophoresis capillary section (3), and a leading liquid electrode tank (4). It is mainly composed of.

The electrophoresis thin tube part (3) has a sample injection port (6) at a bent part (5) in the middle, and a drain port (7) on the leading liquid electrode tank (2) side of the sample injection port. A potential gradient detector (8) is installed on one side of the leading liquid electrode tank (4). Here (9) is the drain port (7)
The pressurized closing valve, 00) is the septum of the sample injection port (6). Note that both the electrode tanks (2) and (4) can be opened to the atmosphere by switching.

Further, to explain in detail the structure around the bending part (5), in FIG. , the needle (12) of the microsyringe (11) is configured to advance through the electrophoresis tube on the leading liquid electrode 11i f'l) side. On the other hand, the drain port is installed on the outer horizontal part of the bent part (5).

Next, the operation of the capillary isotachophoresis analyzer (1) having the above configuration will be explained.

First, in FIG. 2, the terminal electrolyte is introduced from the terminal liquid electrode tank (2) side. In this case, drain port (7
) is released, the terminal electrolyte (T) is sequentially discharged from its drain port (7), but at the point when the terminal electrolyte (T) is discharged, the introduction of the terminal electrolyte is stopped and one of the leading electrolytes (T) is discharged. L) is introduced from the leading liquid electrode tank (4) side. Thus, the leading electrolyte is drained from the drain port (7).
) and is discharged together with some terminal electrolyte.

When the drain port (7) is closed by the on-off valve (9) at the time of this discharge, a boundary (P) between both electrolytes is clearly formed at a predetermined position.

Here, the sample is injected from the sample injection port (6) using the microsyringe (11) as shown in FIG. In other words, the needle (12) of the microsyringe (11) is septum 00).
leading electrolyte (L) in the electrophoresis tube (3).
) is injected only into the area. Therefore, when injecting this sample, the nail (12) does not pass through the terminal electrolyte (T) at all, so a mixing zone of rain and snow dissolution is hardly formed, and separation can be performed in a short electrophoresis time. become.

That is, as described above, after injecting a sample of 3°M and tc, isokinetic electrophoresis is performed by supplying a constant current from a constant current high voltage current source (not shown). ka<b'r sample ion (anion)
are divided into minutes 1i1ff (
Each zone moves at a constant speed in the six directions of arrows with a constant width determined by the amount of ions while maintaining clear boundaries with each other. In this case, a unique potential gradient is formed in each zone depending on its mobility, so this potential gradient is detected by the detector (8) and the separated single component ions are detected. You can know. However, the target substance ions to be fractionated can be detected from the potential gradient value.

In addition, the rain and snow are dissolved in order as described above.
When the drain port (7) is blocked after introducing the liquid into the liquid, if you switch only the leading liquid electrode tank (4) to the atmosphere in advance, the boundary surface (P) for rain and snow dissolution will be at the drain port (7). Since it is almost unaffected by the applied pressure, it can be formed more clearly and in a predetermined position.

Furthermore, when injecting the sample, prepare the terminal liquid electrode tank (2) in advance.
Even if a large amount of test 1'31 is introduced, if only the sample is changed to open to the atmosphere. Since the sample is transferred to the terminal liquid electrode tank (2) and injected, a predetermined migration distance can be ensured and accurate separation analysis can be performed. It also becomes possible to introduce large amounts of samples with low concentrations.

[Brief explanation of the drawing]

Fig. 1 is a functional explanatory diagram showing an example of a capillary type isotachophoresis analyzer for carrying out the present invention, Fig. 2 is an enlarged cross-sectional view around the sample injection port, and Fig. 3 is a diagram showing a conventional example. This is a diagram equivalent to Figure 1. (1)... Capillary type isotachophoresis analyzer, (2
)...Terminal liquid electrode layer, (3)...
・Migration thin tube section, (4)...Leading liquid electrode tank, (6)...Mark test oblique injection ['', (7)...Drain port, (8)... ...Potential gradient detector.

Claims (1)

[Claims] 1. A terminal electrolyte and a leading electrolyte are introduced into the electrophoresis tube from the terminal liquid electrode tank side and the leading liquid electrode tank side, respectively, and both electrolytes are drained with a pressurized on-off valve. The introduction of both electrolytes is stopped when the electrolytes are discharged from the mouth, and then the sample injected into the electrophoresis tube from the sample injection port is subjected to isokinetic electrophoresis to detect the target substance in the sample ■One-tube type In isotachophoresis analysis, the terminal electrolyte is introduced from the terminal liquid electrode tank into the electrophoresis tube, and the electrolyte includes the position of the sample injection port and is located at a drain port located unevenly in the electrophoresis tube on the leading liquid electrode layer side. A capillary isokinetic electrophoresis analysis method characterized in that the introduction of the terminal electrolyte is stopped when the leading solution is discharged from the electrode tank, and after the introduction of the terminal electrolyte is stopped, the terminal electrolyte is introduced from the leading solution electrode tank into the electrophoresis capillary.