JPS635699B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an electrophoresis analyzer, and more particularly, to an electrophoresis analyzer that can easily perform a type of zone electrophoresis in addition to its original function of isokinetic electrophoresis. Regarding.

In isotachophoresis, two types of electrolytes, a terminal liquid and a leading liquid, are filled inside the electrophoresis tube, and substances that become charged at the interface between these two electrolytes, such as amino acids, peptides, biological substances, etc. A sample is injected and subjected to constant current electrophoresis, and the target substance is separated into a single compartment based on the difference in mobility, and qualitatively and/or quantitatively determined using a detector as appropriate. It has become indispensable for analysis.

However, for example, when the amount of the sample is extremely small or when it contains two components with extremely close mobilities, it has not been possible to conduct a sufficient analysis using normal isotachophoresis. That is, in the former case, the width of the ionic component compartment of the obtained sample is too narrow for sufficient detection, and in the latter case, the ionic component compartments of the two components are too close to each other to distinguish them. Something happened that I couldn't do.

The present invention provides an improved electrophoresis analyzer that can suitably perform analysis even in the above-mentioned cases. In an electrophoresis analyzer in which a sample injection part and a detector are connected in order between a liquid electrode tank and a leading liquid electrode tank by a pipe line, a sample injection part is installed in an arbitrary part of the pipe line from the sample injection part to the leading liquid electrode tank side. a switching connection section, an arrival detection means for detecting that an ion component compartment has arrived at the switching connection section, and a sample injection section for zone electrophoresis analysis, and the switching connection section connects the electrophoresis connection pipe and the zone. and a connecting pipe for injecting an electrophoretic solution for electrophoresis, and by switching, any one of the connecting pipes is connected to the pipe, and by another switch, the connecting pipe for electrophoresis is connected to the sample for zone electrophoresis analysis. An electrophoretic analysis device is provided that is connected to an injection section.

According to the apparatus of the present invention, ordinary isotachophoresis and zone electrophoresis can be easily linked, so that even samples that cannot be sufficiently analyzed by ordinary isotachophoresis alone can be suitably analyzed. Furthermore, since standard samples can be subjected to zone electrophoresis under the same conditions, direct comparative analysis becomes possible.

Hereinafter, the present invention will be explained in further detail based on embodiments shown in the drawings. However, this invention is not limited thereby.

Reference numeral 1 shown in FIG. 1 is an embodiment of an electrophoretic analyzer according to the present invention. A terminal liquid electrode tank 3 and a leading liquid electrode tank 4 are connected to both ends of the high voltage power supply circuit 2, and a sample injection unit 5 is connected between these.
and a detector 6 are connected through a conduit 7 to constitute a basic analysis section.

Terminal liquid electrode tank 3 is connected to terminal liquid inlet 8
and terminal liquid tank 10 via valve 9
The leading liquid electrode tank 4 is connected to a leading liquid tank 13 via a leading liquid inlet 11 and a valve 12. The terminal liquid inlet 8 is further connected to an electrolyte tank 15 for zone electrophoresis via a valve 13, and the leading liquid inlet 11 is similarly connected to an electrolyte tank 15' for zone electrophoresis via a valve 13'. ing.

The central part of the conduit 7 between the sample injection part 5 and the detector 6 has been removed, and a switching connection 30 is provided in the removed part. That is, the removed ends of the pipe line 7 are respectively formed into sliding flanges 16 and 16', and a sliding body 17 is slidably interposed between the sliding flanges 16 and 16'. A connecting pipe 18 for migration and a connecting pipe 18' for zone electrophoresis are bored in the sliding body 17, and by sliding the sliding body 17, the connecting pipe 7 can be connected to any of the connecting pipes. It is possible to connect the removal parts of Further, a second sample injection section 19 separate from the sample injection section 5 is provided in a part of the sliding flange 17, and a sample injection channel 25 is connected to the sample injection section 19. By sliding the sliding body 17,
Connecting conduit 18 can be connected to second sample injection section 19 .

The electrophoresis current supplied from the high voltage power supply circuit 2 to the analysis section is input to the microcomputer 22 via the current detection circuit 21. These current detection circuit 21 and microcomputer 22 constitute an arrival detection means, and detect to which position in the conduit 7 the ion component section of the sample has electrophoresed. That is, as is well known, the amount of electricity supplied during electrophoresis is directly proportional to the migration distance of one ionic component compartment. Therefore, by knowing the amount of electricity, the migration distance can be determined. The microcomputer 22 integrates the output of the current detection circuit 21, that is, the electrophoretic current, to obtain the above-mentioned amount of electricity, and thereby detects the electrophoretic position of the ion component section.
Now, if we pay attention to the ion component compartment of the leading liquid, the relationship between the migration distance and the amount of electricity depends only on the composition of the leading liquid. However, since the rear end of the ionic component compartment of the leading solution is nothing but the front end of the ionic component compartment of the sample, the migration distance of the front end of the ionic component compartment of the sample also depends on the relationship between the migration distance of the ionic component compartment of the leading solution and the amount of electricity. You can know it. In other words, as long as the leading liquid is the same, the migration distance of the ionic component compartment of the sample can be determined from the electrical quantity using the same judgment conditions, and there is no need to consider the composition or amount of the sample. . For this purpose, in this apparatus 1, the composition of the leading liquid and the amount of electricity required for the rear end of the ion component compartment of the leading liquid to migrate from the sample injection part 5 to the connecting tube outlet 20 when the leading liquid is used are determined in advance. are stored in the microcomputer 22 in association with each other.

23 is a drive mechanism for the sliding body 17, which is controlled by the microcomputer 22.
Therefore, the microcomputer 22 and the drive mechanism 23 constitute means for selecting and switching the connecting pipes 18 and 18'.

24 is a console for interacting with the microcomputer 22.

Next, the operation of this device 1 will be described, assuming that the removal portion of the pipe line 7 is connected by the connecting pipe line 18 as an initial state. First, the composition of the reading liquid is input into the microcomputer 22 via the console 24. Then, the microcomputer 22 reads out the corresponding quantity of electricity Q from the memory and sets it internally. Next, the drain stock 26 and valve 12 are opened, and the leading liquid is filled in the pipe line 7 on the leading liquid electrode tank 4 side from the sample injection part 5. Furthermore, the valve 9 is opened and the pipe line 7 on the opposite side is filled with the terminal liquid. , close each valve. In this way, an interface between the terminal liquid and the leading liquid is created in the sample injection unit 5, a sample is injected into the interface, and a link analysis start command is input from the operation console 24. Upon receiving the link analysis start command, the microcomputer 22 controls the power supply circuit 2 to output a migration current and starts isokinetic electrophoresis.
The electrophoresis current i ₁ is input to the microcomputer 22 by the current detection circuit 21, and the microcomputer 2
2 integrates the electrophoretic current i ₁ over time and compares the amount of electricity q obtained by the integration with the amount of electricity Q. FIG. 1 and FIG. 2a show this state, in which the ionic component compartment is electrophoresed at a constant velocity in the direction of the arrow α.

When the rear end of the ionic component section of the leading liquid reaches the connecting pipe outlet 20, the integrated quantity of electricity q matches the quantity of electricity Q. Therefore, the computer 22 operates the power supply circuit 2 to stop the electrophoresis current, and also operates the drive mechanism 23 to disconnect the connecting pipe 18 from the electrophoretic pipe 7 and connect it instead. Conduit 18' is connected to migration conduit 7. As a result, the ionic component section of the sample that follows the leading liquid ionic component section is trapped in the connecting conduit 18. At this point, the computer 22 notifies the operator via the console 24 that preparations for injection of the electrolytic solution for zone electrophoresis are complete.

The operator opens the drain tank 26 to drain all the electrolyte in the pipe line 7, and then opens the valves 13 and 1.
3' is opened to fill the conduit 7 with an electrolytic solution for zone electrophoresis, and after closing the valves 26, 13, and 13', a command to start zone electrophoresis is inputted from the console 24.

This causes the computer 22 to control the drive mechanism 23.
is activated, and the connection pipe 1 is opened again as shown in Fig. 2a.
8 is connected to the conduit 7, and the power supply circuit 2 is activated to output electrophoresis current _i2 . The ionic component compartments of the sample that were trapped in the electrophoresis tube 18 begin electrophoresis again, but this is zone electrophoresis.

In the end, the sample is subjected to isokinetic electrophoresis from the sample injection section 5 to the removal section, and zone electrophoresis from the removal section to the detector 6, that is, link electrophoresis, and is detected.

When analyzing a standard sample, first, a standard analysis start command is input from the console 24 to the microcomputer 22. Microcomputer 2
2 operates the drive mechanism 23 to set the switching connection part 30 to the position shown in FIG.

Therefore, the operator points the arrow β to the sample injection section 25.
After flowing the standard sample as shown below to fill the connecting pipe 18 with the standard sample, a standard sample injection command is input from the operation console 24.

Then, the microcomputer 22 operates the drive mechanism 23 to set the switching connection part 30 to the position shown in FIG.

The subsequent operation is the same as the latter half of the link analysis, so the explanation will be omitted, but in short, only the zone electrophoresis of the standard sample can be easily performed.

Modified embodiments include one in which the switching connection part 30 is of a rotating disk type, one in which the number of connection pipes 18, 18' is further increased, each valve 9, 12, 13, 13',
26 is a solenoid valve and the microcomputer 22
A type in which only a specific ion component section of the sample is trapped and subjected to zone electrophoresis, and a type in which both electrode tanks 3 and 4 are automatically controlled.
one that uses a reach detection means that detects the arrival position of the ionic component compartment based on the temporal change in the voltage between;
An example is one in which the switching connection part 30 is provided in the conduit 7 between the detector 6 and the leading liquid electrode tank 4.

Note that the pipe line 7 from the sample injection section 5 to the removal section
However, if the capacity is sufficiently larger than that of the conduit 7 from the removal section to the detection section 6, the electrolytic solution for zone electrophoresis can be transferred from the sample injection section 5 to the conduit 7 on the leading liquid electrode tank 4 side. Zone electrophoresis may be performed with only the electrolyte tank 13 filled with the electrolyte tank 15. In this case, the valve 13 and the electrolyte tank 15 become unnecessary. Furthermore, if the electrolytic solution for zone electrophoresis is used as a leading solution, 13',
15' is also unnecessary.

As understood from the above description, according to the electrophoresis analyzer of the present invention, isotachophoresis and zone electrophoresis can be easily linked.
Furthermore, since it is possible to perform only isokinetic electrophoresis without switching the connecting pipes, there is an advantage that the range of samples that can be analyzed can be greatly expanded. Furthermore, since the standard sample can be injected into the position where zone electrophoresis is started during link analysis and zone electrophoresis is performed, there is also the advantage that comparative analysis can be performed directly.

[Brief explanation of the drawing]

FIG. 1 is a diagram illustrating the configuration of an embodiment of the electrophoretic analysis device of the present invention, and FIG. 2 is a diagram illustrating each switching position of the connecting channel of the device shown in FIG. 1. DESCRIPTION OF SYMBOLS 1... Electrophoresis analyzer, 2... High voltage power supply circuit, 3... Terminal liquid electrode tank, 4... Leading liquid electrode tank, 5... Sample injection part, 6... Detector, 7... Pipe line , 16... sliding flange, 17...
...Sliding body, 18, 18'...Connection pipe, 19...
Sample injection unit for zone electrophoresis analysis, 21...Current detection circuit, 22...Microcomputer, 23
...Drive mechanism.

Claims (1)

[Scope of Claims] 1. An electrophoresis analyzer in which a sample injection part and a detector are connected in order by a pipe between a terminal liquid electrode tank and a leading liquid electrode tank, which are respectively connected to both ends of a high voltage power supply circuit. A switching connection section provided at an arbitrary part of the conduit from the sample injection section to the leading liquid electrode tank side, an arrival detection means for detecting that an ion component compartment has reached the switching connection section, and a device for zone electrophoresis analysis. The switching connection part has a connecting pipe for electrophoresis and a connecting pipe for injecting an electrolytic solution for zone electrophoresis, and any one of the connecting pipes can be connected to the pipe by switching, and the connecting pipe can be connected to the other by switching. An electrophoresis analysis apparatus characterized in that the electrophoresis connecting pipe is connected to the zone electrophoresis analysis sample injection section by switching. 2. The device according to claim 1, wherein the switching connection portion is provided at a central position of the conduit between the sample injection portion and the detector. 3. The switching connection part is composed of sliding flanges provided at both removal ends of the pipe line, and a sliding body having two connecting pipe lines and provided between the sliding flanges and slidingly moving, 3. The apparatus according to claim 1, wherein a sample injection part for zone electrophoresis analysis is provided on the sliding flange. 4. Claims 1 to 3, wherein the arrival detection means comprises a migration current detection circuit, an integration circuit for the output of the migration current detection circuit, and a comparison circuit that compares the output of the integration circuit with a predetermined threshold value. Apparatus according to any of paragraphs.