JPS58148951A - Analyzing apparatus of electrophoresis - Google Patents

Abstract

PURPOSE:To perform a sufficient analysis even with a trace sample, by connecting one optional switch connection part to a duct by switching and also, connecting a connection duct for migration to a sample injection part for a zone electrophoresis analysis by another switching. CONSTITUTION:A removal part of a duct 7 is connected by a connection duct 18 as an early condition. At first, when the composition of a leading liquid is inputted to a microcomputer 22 through a console desk 24, the quantity of electricity Q corresponding to the microcomputer 22 is read out from a memory by the microcomputer 22 and is set internally. Next, a drain cock 26 and a valve 12 are opened and the leading liquid is filled up the duct 7 of a leading liquid electrode cell 4 side from a sample injection part 5. Moreover, a terminal liquid is filled up the duct 7 by opening a valve 9 and each valve is shut. In this manner, the boundary face is formed between the terminal liquid and the leading liquid at the part 5 and the sample is injected to the boundary face. Then, the analysis is performed by a link analysis start command from the desk 24.

Description

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

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 types of pI# liquids, such as amino acids, peptides, and living organisms, are 9! Electrophoresis is performed using a constant current by injecting a sample such as J quality, and the target substance is separated into a single compartment by the difference in mobility, and the substance is analyzed and/or quantified using a detector as appropriate. It has become indispensable for sample analysis.

However, for example, when the sample is in an extremely small amount or contains components with very close mobilities of 2 fs, optical analysis cannot be performed using normal isotachophoresis. That is,
In both cases, the ionic component zones of the resulting sample are too wide to allow sufficient detection, and in the latter case, the ionic component zones of the two components are too close to distinguish between them. Something like that happened.

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 by a pipe between a terminal liquid electrode tank and a leading liquid electrode tank, any part of the pipe from the sample injection part to the leading liquid electrode side a switching connection section provided in the switching connection section, an arrival detection means for detecting that the ion component compartment has reached the switching connection section, and a sample injection section for zone electrophoresis analysis; and a connecting conduit for injecting an electrolyte for zone electrophoresis, and by switching, any one of the connecting conduits is connected to the conduit, and by the other switching, the connecting conduit for primary 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, it is possible to link normal isotachophoresis and zone electrophoresis to a container, so that it is possible to suitably analyze samples that cannot be analyzed sufficiently with ordinary isotachophoresis alone. become. In addition, since the standard sample can be subjected to zone electrophoresis under Oka conditions, comparative analysis can be performed in a direct manner.

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

The ill shown in FIG. 1111 is an embodiment of the electrophoresis analyzer of the present invention. A terminal liquid electrode tank (3) and a leading electrode tank (4) are connected to both ends of the voltage power supply port * (2+), and a sample injection part (5) is connected between them.
and a detector (6) are connected by a tube M fi+ to constitute a basic analysis section.

The terminal liquid electrode tank (3) is connected to the terminal liquid inlet (8).
The leading liquid electrode tank (4) is connected to the leap ink liquid tank Q3 via the leading liquid inlet l and the valve @. The terminal liquid inlet (8) is further connected to the electrolyte tank for zone electrophoresis via the valve Ω.
Similarly, the leading liquid inlet a' is also connected to the zone electrophoresis electrolyte tank (15) via the valve (13).
').

The central part of the conduit (7) between the sample injection part (5) and the detector (6) has been removed, and the removed part has a switching connection ■
is provided. That is, the removal end of the conduit (7) is
A sliding body α is formed between the two sliding flanges α and G (Iff), respectively.
η is slidably interposed. The sliding body αη has a connection pipe a for electrophoresis and a connection pipe for zone electrophoresis (1g
The removed part of the pipe (7) can be connected to any of the connecting pipes by sliding the sliding corporal punishment. Another sliding 7 lunge Q! A second sample injection section (2) separate from the sample injection section (5) is provided in a part of l, and a sample injection section (a) is connected to the sample injection section α. . By sliding the sliding body surface, the connecting tube w1 (to) can be connected to the second sample injection section 4.

The electrophoresis current supplied from the high voltage power supply circuit (2) to the analysis section is transmitted via the current detection circuit 0 to the microcomputer@
is man-powered. These current detection circuit (to) and microcomputer (to) constitute an arrival detection means, and detect to which position in the conduit (7) the ionic 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 ion component section. Therefore, by knowing the amount of electricity, the migration distance can be determined. The microcomputer @ integrates the output of the current output circuit 1I11@, that is, the electrophoretic current, to obtain the above-mentioned quantity of electricity, and thereby detects the electrophoretic position of the ion component section. Now, if we focus on the ionic component compartment of the leading liquid, the relationship between the migration distance and the amount of electricity is determined only by the composition of the leading liquid. However, since the rear end of the ionic component section of the leading liquid is nothing but the front end of the ionic component section of the sample, the migration distance of one end of the ionic component section of the sample is also the same as the electrophoretic distance of the ionic component section of the leading liquid and the amount of electricity. It can be known through relationships. In other words, since the leading liquid is the same and the same judgment conditions are used, the migration distance of the ionic component compartment of the sample can be determined from the electrical quantity, and there is no need to consider the composition of the sample or the lid. Become. For this purpose, in advance, in this apparatus 1tll, we have determined the composition of the leading liquid and the migration of the rear end of the ion component compartment of the leading liquid from the sample injection part (5) to the connecting tube outlet (to) when the leading liquid is used. The amount of electricity required for is stored in the microcomputer (2a4c) in association with each other.

(to) is a drive mechanism for the sliding body αη, which is controlled by a microcomputer (2). Therefore, the microcomputer @ and the drive mechanism are connected to the connecting pipe u (L
8ri0) It constitutes a selection switching means.

(Foundation) is a console for interacting with a microcomputer.

Next, the operation of this device 11 will be explained, assuming that in the initial state, the removal section of the pipe line (7) is connected to the connecting pipe line (to). First, the composition of the leading liquid is input into the microcomputer (2) via the operation console (to).

When the current is generated, the microcomputer extracts the corresponding quantity of electricity Q from memory and sets it internally.
. Next, open the drain cock (to) and the valve, and pour the leading liquid from the sample injection part 4 into the leading liquid electrode tank (
4) Fill the pipe line (η) on the side, then flip the valve (91) to fill the pipe line (7) on the opposite side with the terminal liquid, and close each valve.In this way, the boundary between the terminal liquid and the leading liquid is placed in the sample injection part t51. A surface is created, a sample is injected into the boundary surface, and a link analysis start command is input from the operating vehicle (to).When the microcomputer receives the link analysis start command, it controls the electrophoresis circuit (11). A current is output and isokinetic electrophoresis is started.The electrophoresis current 51 is input to the current detection circuit (to) the microcomputer, and the microcomputer (2) detects the electrophoresis type fiit.
is integrated over time, and the electrical quantity obtained by the integration is compared with the electrical quantity Q. Figures 1 and 2
Figure (1) shows this state, in which the ionic component compartment undergoes constant electrophoresis in the direction of the arrow -.

The rear end of the ionic component compartment of the leading liquid is the connecting pipe outlet.
When it reaches , the integrated electricity meter is equal to the quantity of electricity Q by 1k. Therefore, the computer @ operates the power supply circuit M121 to stop the electrophoresis current, and also operates the drive mechanism (to) to connect the connecting conduit (to) to the electrophoresis tube as shown in FIG. 2(b). (7), and connect the connecting pipe (1B') to the electrophoresis pipe (7) instead. As a result, the ionic component compartment of the sample that follows the leading liquid ionic component compartment is trapped in the connecting pipe (to).

At this point, the computer notifies the operator via the console that the preparation for filling the electrolytic moat for zone electrophoresis is complete.

The operator opens the drain cock (to) and drains pipe II (7).
Drain all the electrolyte in 1, then close the valve (13 (If
After filling the pipe (7) with the electrolytic solution for zone electrophoresis and closing the valve 13 (Iff), enter the zone electrophoresis star command manually from the console.

As a result, the computer operates the drive mechanism (2) to again connect the connecting pipe (71) to the pipe (71) as shown in Figure 2 (a), and operates the power source (1w1 (2)) to output the electrophoretic wire (i). The ionic component compartments of the sample that were trapped in the migration tube (2) start electrophoresis again, or this is zone electrophoresis.

As a result, the sample is subjected to isokinetic electrophoresis from the sample injection section (51) to the removal section, and is subjected to zone electrophoresis from the removal section to the detector (6), that is, link electrophoresis, and is detected.

Now, when analyzing a standard sample, first, a standard analysis start command is manually input from the operating vehicle □ to the microcomputer @. The microcomputer 嬬 operates the drive #J@structure, sets the switching connection part (to) to the position shown in Fig. s2 (C), and places the operation console (
) notifies the operator that preparation for specimen injection is complete.

Therefore, the operator points the arrow (branch) into the sample injection channel (2).
After pouring the standard sample into the throat and filling the connecting pipe (layer) with the standard sample, enter the standard injection completion command from the operation console ■.

The controller and microcomputer operate the drive mechanism a to set the switching connection part (■) to the position shown in Figure 2 (b), and the operator via the console (to) tells the operator that the preparation for zone electrophoresis is complete. Notify.

This excellent operation is the same as the latter half of the link analysis mentioned above, so the explanation will be omitted, or in short, the zone 1 of the standard sample.
Pneumophoresis alone can be easily performed.

Modified embodiments include one in which the switching connecting part circle is a rotating disk type, one in which the number of connecting pipes TNI (to) (1 ") is further increased, and each valve +91 (12 (13 (1: Y) Cal) is replaced with a solenoid valve. One that is automatically controlled by a microcomputer @, one that traps only a specific ion component section of the sample and performs zone electrophoresis, and one that traps only a specific ion component section of the sample and performs zone electrophoresis. A device using a reaching detection means for detecting the reached position of the ion component compartment when the ion component compartment arrives, and a device in which a switching connection (to) is provided in the pipe m1 (71) between the detector 1 tin and the leading liquid electrode tank (4). Examples include.

Note that if the pipe line (7) from the sample injection part (5) to the removal part has a sufficiently larger capacity than the pipe line (7) from the removal part to the detection part (6), the zone As can be understood from the above explanation, zone electrophoresis is performed with the electrolytic solution for electrophoresis filled from the sample injection part (5) to only the leading liquid electrode tank (pipe line (7) on the a side). According to the electrophoresis analyzer, it is possible to link isotachophoresis and zone electrophoresis in a container, and it is also possible to perform only isotachophoresis without switching the connecting pipe. This has the advantage of greatly expanding the range of samples that can be analyzed.Furthermore, during link analysis, a standard sample can be injected at the starting position of zone electrophoresis and zone electrophoresis can be performed, making specific soft analysis possible. There are also advantages to doing it directly.

[Brief explanation of drawings]

FIG. 1 is a diagram illustrating the configuration of an embodiment of the electrophoretic analysis device of the present invention, and FIG. 182 is a diagram illustrating each switching position of the connecting channel of the device shown in FIG. 1. (1)...Par electrophoresis analyzer, (2)...High voltage power supply circuit, (3)...Terminal liquid electrode, (4)...
・Leading liquid electrode tank, (5)...Sample injection part, (
6)...Detector, (7)...Pipe line, West...Sliding flange, αη...Sliding body, (li9 (Ig)...
Connecting pipe, sample injection part for zone electrophoresis analysis,
(to)... ill inflow/output circuit ■... microcomputer, two... drive mechanism.

Claims (1)

[Scope of Claims] 1. Electrophoretic analysis in which a sample injection part and a detector are sequentially connected by a conduit between a terminal liquid electrode and a leading liquid electrode which are respectively connected to both ends of a high voltage power supply circuit. In the apparatus, a switching connection part provided at an arbitrary part of the conduit from the sample injection part to the leading liquid electrode side, an arrival detection means for detecting that an ion component compartment has reached the switching connection part, and a zone electrophoresis analysis device. The primary switching connection part has a connecting pipe for electrophoresis and a connecting pipe for injecting electrolyte for zone electrophoresis, and any number of 10,000 can be connected to the first pipe by switching. Furthermore, the electrophoresis analyzer is characterized in that the electrophoresis connecting pipe is connected to the zone 1 electrophoresis analysis sample injection part by another switch. 2. The device according to claim 1, wherein the switching connection portion is provided at a central position tmti of the conduit between the sample injection portion and the detector. 3. The switching connection part is composed of sliding 7 langes provided at both removal ends of the conduit, and a sliding body having two connecting conduits and provided between the 7 lunges. The apparatus according to claim 1 or 2, wherein the sample injection part for zone electric vibration analysis is provided on the sliding 7 flange. 4. The arrival detection means comprises a migration current detection path, an integration circuit for the output of the migration current detection circuit, and a ratio circuit for comparing the output of the integration circuit with a predetermined threshold value. The device according to any one of items 1 to 3.