JPS6257216B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an electrophoretic analysis method and apparatus, and more particularly, to an electrophoretic analysis method and apparatus in which an injected sample or its target component is repeatedly electrophoresed many times without taking it out of the system. Regarding.

Conventionally, when performing high separation of samples by electrophoresis, the length of the electrophoresis tube was increased, counterflow was used, or the sample components separated by electrophoresis were taken out of the carrier system and reinjected. I used to repeat electrophoresis.

However, those that increase the length of the electrophoresis tube have the disadvantage that the overall Joule heat increases and bubbles are more likely to be generated, and those that use a counterflow have the disadvantage that the electrolyte flows into the ionic component zone. It has the disadvantage of causing turbulence, and those that separate it out of the carrier system have the disadvantage that the operation is troublesome and a part of the sample may be lost during separation.

This invention was made with the aim of solving the above-mentioned drawbacks. Specifically, an electrophoresis tube and a trap section are arranged on both sides of the detector, and the injected sample is electrophoresed. The target component that has passed through the detector and been separated by electrophoresis is trapped in the trap section in the migration direction, and the electrolyte is exchanged and the applied voltage is switched to transfer the target component to the detector in the opposite direction. to be electrophoresed through the trap, and to be trapped in another trap section.
An electrophoretic analysis method in which a target component is electrophoresed alternately in both directions across a detector to precisely separate and analyze the target component, and an apparatus suitable for carrying out the method, that is, a high-voltage power supply circuit A first trap is connected between the electrode tanks connected to both ends of the
A drain switching means, a detector, and a second trap-drain switching means are sequentially connected by a pipe line, and a pipe line between the first trap-drain switching means and the detector, and a pipe line between the first trap-drain switching means and the detector, and a pipe line between the first trap-drain switching means and the detector, The conduit between the two trap-drain switching means is constituted by a separation migration tube, and the first and second trap-drain switching means are connected to both ends of each at their first positions. A corresponding trap pipe is inserted between the electrode tank and the detector, and a corresponding drain is inserted in place of the trap pipe at the second position, and the first and second trap pipes
At least one of the drain switching means is configured to connect the corresponding trap tube to the sample inlet in the second position.

The present invention is effective for precisely separating a target component from a variety of coexisting charged substances, for example, in the analysis of charged substances in foods or biological samples.

Hereinafter, this invention will be explained in detail based on embodiments shown in the drawings. Note that this invention is not limited to this.

1 shown in FIG. 1 is an embodiment of an electrophoretic spectrometer according to the present invention. A first electrode tank 3 is connected to one end of the high voltage power supply circuit 2, and a second electrode tank 4 is connected to the other end.
The Happo Kotsukku 5, the detector 6, and the second Happo Kotsukku 7 are connected in order through conduits 8, 9, 10, and 11.

The detector 6 consists of a pair of potential gradient detectors juxtaposed with a short distance apart in the electrophoresis direction,
Detection of migration speed is also possible.

Pipe lines 9 and 10 are capillary reach tubes for separation.

When the first Happo pot 5 is switched to the first position (the flow path position in the pot shown by the solid line in FIG. 1), the trap pipe 12 is inserted between the conduits 8 and 9, (Flow path position in the pot indicated by the broken line in Figure 1)
When the switch is switched to , a drain 15 is inserted in place of the trap pipe 12 via the drain stocks 13 and 14. Furthermore, in this second position, one end of the trap tube 12 is connected to the sample injection port 16, and the other end is connected to the drain 15 via the drain tank 17.

In the second Happo stock 7, the trap pipe 18, drain stock 19, 2
0, 23, a drain 21, and a sample injection port 22 are connected.

When performing the analysis, first move the first Happo Kotoku 5 to the second position (broken line), and move the second Happo Kotoku 7 to the first position.
Set the drain to the position (solid line), and
14, the terminal electrolyte T ₁ is filled between the first electrode tank 3 and the first Happo pot 5, and the leading electrolyte is filled between the second electrode tank 4 and the first Happo pot 5. Fill with liquid L ₁ . Furthermore, while operating the drain stock 17, the sample S is injected from the sample injection port 16, and the trap tube 12 is
Fill the inside with sample S.

Here, if the first Happo pot 5 is switched to the first position (solid line) and a constant current is supplied from the high voltage power supply circuit 2, the sample S starts electrophoresing in the direction of the detector 6.

For example, the detector 6 obtains outputs as shown in FIG. 2, and if B is the target component, then
Since the time when the target component B enters the trap tube 18 can be estimated based on the migration speed obtained by the detector 6 and the known distance to the trap tube 18, the second Happo pot 7 is moved to the second position ( (dashed line) and stop the current at the same time. At this time, the sample injection port 22 and the drain tank 23 are kept closed.

Next, the drain tanks 19 and 20 are opened, and all the electrolyte other than the part collected in the trap pipe 18 is discharged into the drain 21. Then, in the same manner as previously done in the first Happo pot 5, a new leading electrolyte _L2 is filled between the first electrode tank 3 and the second Happo pot 7, and the second electrode tank is filled with a new leading electrolyte L2. 4
A new terminal electrolyte T ₂ is filled between the time and the second Happo Kotoku 7.

After that, if the second Happo pot 7 is returned to the first position (solid line) and the polarity of the high voltage power supply circuit 2 is reversed to flow a constant current in the opposite direction, the target component collected in the trap tube 18 B performs electrophoresis in the opposite direction to the previous one. Note that at this time, since the signal direction at the detector 6 is reversed, it is preferable to perform appropriate adjustment.

Thereafter, by repeating electrophoresis between the first Happo-kottoku 5 and the second Happo-kottoku 7 while monitoring the separation state with the detector 6, the target component B can be separated more precisely, e.g. At the m-th time, highly separated outputs as shown in FIG. 3 are obtained.

As is clear from the above description, according to the electrophoretic analysis method and apparatus of the present invention, it is possible to easily perform repeated electrophoresis of an injected sample or its target components many times without taking them out of the system. Furthermore, since it is possible to easily change the analysis conditions such as changing the composition of the electrolytic solution, the sample can be highly separated very suitably and reliably.

Another advantage is that if bubbles occur in the electrophoresis tube, they can be expelled each time the electrophoresis direction is switched.

Note that the present invention can also be applied to electrophoresis that uses a single electrolytic solution without using a leading electrolytic solution and a terminal electrolytic solution. Further, since at least one trap-drain switching means only needs to be connected to the sample injection port, the other trap-drain switching means may be, for example, a hexagonal socket 7' as shown in FIG. 4.

[Brief explanation of the drawing]

Fig. 1 is an explanatory diagram of the configuration of an embodiment of the electrophoretic spectrometer of the present invention, Fig. 2 is an output diagram of the detector during the first electrophoresis, and Fig. 3 is the detector output during the m-th electrophoresis. FIG. 4 is an explanatory diagram of a configuration of a modified example of the trap-drain switching means. 1... Electrophoretic spectrometer, 2... High voltage power supply circuit, 3... First electrode tank, 4... Second electrode tank,
5...First Happo Kotoku, 6...Detector, 7...
Second Happo Kotoku, 8, 9, 10, 11... Pipeline, 9, 10... Capillary reach tube, 12,
18, 18'... Trap tube, 13, 14, 1
7, 19, 20, 23, 19', 20'...Drain, 15, 21, 21'...Drain, 16,
22...Sample injection port.

Claims (1)

[Scope of Claims] 1. Purpose of electrophoresis separation after passing through the detector using a device that has an electrophoresis tube and a trap section on both sides of the detector, and separates the injected sample by electrophoresis. The components are trapped in the trap section in the migration direction, and the electrolyte is replaced and the applied voltage is changed to cause the target component to electrophores in the opposite direction through the detector, and then to be trapped in another trap section. , an electrophoretic analysis method characterized in that target components are electrophoresed alternately in both directions across a detector, and the target components are precisely separated and fractionated. 2. A first trap-drain switching means, a detector, and a second trap-drain switching means are sequentially connected by a conduit between the electrode tanks respectively connected to both ends of the high voltage power supply circuit, and the A conduit between the first trap-drain switching means and the detector and a conduit between the detector and the second trap-drain switching means are constituted by a separation electrophoresis tube, In the trap-drain switching means, in the first position, a corresponding trap tube is inserted between the electrode tank and the detector connected to each end, and in the second position, the trap tube is replaced with the trap tube. The trap tube is configured to insert a corresponding drain, and at least one of the first and second trap-drain switching means is configured to connect the corresponding trap tube to the sample inlet in the second position. An electrophoretic spectrometer.