JPS58130Y2 - Isokinetic electrophoresis analyzer - Google Patents

Description

[Detailed explanation of the idea] This invention relates to an isotachophoresis analyzer.

More specifically, this invention is an isotachophoresis analyzer equipped with a terminal electrolyte tank, an electrophoresis tube in which a sample injection port and a detector are installed, and a leading electrolyte tank in order. A separation means is provided in the migration pipe a distance A away from the tank side, and an electrolyte pushing means is provided in the migration pipe or the leading electrolyte tank between the detector and the leading electrolyte tank, and the purpose of the detector is Based on the first detection of the substance, the electrolyte pushing means is activated to push the target substance back toward the sample injection port by a distance B from the detector to continue electrophoresis, and after a time T after receiving the first detection signal. The present invention relates to an isotachophoresis analyzer comprising automatic control means for controlling the fractionation means to operate based on a re-detection signal after a time (A/B)T after receiving the re-detection signal.

In isotachophoresis analysis, a terminal electrolyte and a leading electrolyte are filled in a migration tube with a fixed diameter, such as a capillary tube, and charged substances (amino acids, peptides, etc.) are filled at the interface between them. In this method, a sample of biological materials, etc.) is placed, and isokinetic electrophoresis is performed using a constant current to separate the substance to be detected for qualitative and/or quantitative analysis.

However, although the quality is determined by comparing it with a standard, it cannot be determined if the standard is not available, so the zone where the analyte has been separated is usually sampled and qualitative and/or quantitative analysis is performed separately. I was going.

Examples of devices that specifically carry out such fractionation include:
As disclosed in Japanese Patent Laid-Open No. 50-81397, an apparatus is known in which a branch pipe is provided to separate a sample, and a leading electrolyte is forced to flow into the branch pipe to collect the separated sample.

However, in general, the migration speed of the target substance in isotachophoresis analysis is extremely slow, and even if the distance between the detector position and the preparative collection position is made close as in the above-mentioned Japanese Patent Application Laid-Open No. 50-81397, It takes a considerable amount of time from when a substance is detected by a detector until it reaches a collection position, and therefore, there are cases where the target substance cannot be collected accurately.

This invention was devised to solve these problems, and one of its features is that at the same time as the target substance is detected, the target substance is pushed back a certain distance, made to migrate again, and redetected. Another feature is that the time for the target substance to reach the collection position, that is, the time for collection, is determined based on the migration speed determined by redetection.

The specific structural features of this invention and its advantages will become clear from the following description.

This invention will be explained in detail below based on the embodiments shown in the figures.

Note that this invention is not limited by this.

In FIG. 1, a capillary type isotachophoresis analyzer 1 has a constant inner diameter (for example, It mainly consists of a capillary tube 6, a leading electrolyte tank 7, and an automatic control device 8.

The electrolyte pushing/separating means 5 of the electrophoresis tube 6 is connected to a syringe pump 9 for pushing/separating, a septum 10 of the electrophoresis tube 6, and a device for passing through the septum from inside the electrophoresis tube to the syringe pump 9. The opening 12 in the electrophoresis tube 6 of the branch tube 11 is cut at an angle so that the opening direction is on the detector 4 side.

The pushing/separating syringe pump 9 can forcibly push a certain amount of the leading electrolyte toward the detector 4 of the migration tube 6.

The automatic control device 8 includes an amplifier for the detection signal of the target substance by the potential gradient detector 4, a relay, a power supply circuit for the syringe pump 9 described above, a switch, and other electric circuits.

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

First, the interface between an electrolyte containing anions with higher mobility than the sample (leading electrolyte) and an electrolyte containing anions with lower mobility (terminal electrolyte) is inserted into the sample injection port 3 of the electrophoresis tube 6. A sample is injected into the interface, and a constant current is supplied from a constant current high voltage power supply (not shown) to perform electrophoresis (
constant velocity).

In this way, the sample ions (anions) are separated into zones (bands) of single component ions in the electrophoresis tube in order of their mobility, and each zone maintains a clear boundary with each other, while each zone has a constant density determined by the amount of ions. Start moving at a constant speed in the direction of the arrow with a width of .

In this case, different unique potential gradients are formed in the base zone depending on the mobility, and this potential gradient is detected by the detector 4.
Single component ions detected and separated can be known.

That is, target substance ions can be detected from the potential gradient value.

When the detector 4 detects the target substance ion, that is, the boundary surface of this ion, the detection signal is transmitted to the automatic controller 9 at that moment, and the electrolyte pushing/separating means 5 is activated based on the signal. The syringe pump 9 is actuated to force a certain amount of the leading electrolyte through the opening 12 toward the detector 4, thereby pushing the interface of the target substance ions back to a distance B from the detector 4. Electrophoresis (continue at constant speed).

Thus, the detector 4 receives the first detection signal at a time T
Later, when the target substance ion (interface) is re-detected and the re-detection signal is transmitted to the automatic control device 8, the electrolyte is pushed in and out after a time (A/B) T after receiving the re-detection signal.
The syringe pump 9 of the fractionation means 5 is activated to fractionate the target substance.

In other words, the target substance ions have traveled a distance B in a time T, and can be considered to continue moving at the same speed even after re-detection, so they must be moved a distance A to the separation position in a time (A/B)T. do.

Therefore, by performing the fractionation operation at that time, the target substance can be accurately fractionated.

Further, since the target substance is directly collected into the syringe pump 9, even if the amount is small, a large amount can be collected by increasing the number of times the target substance is collected, thereby increasing the accuracy of analysis.

Different from the above embodiments, an electrolyte pushing/separating means may be provided separately.

That is, the electrolyte pushing means can be provided at a position different from the separation position (in the migration conduit or the leading electrolyte tank).

In addition, the preparative separation means divides the electrophoresis narrow channel, and forms flanges on the opposite end faces of the separation, and a preparative plate (rotating or reciprocating) is slidably interposed between these flanges. The plate may be provided with at least one communication hole having the same diameter as the migration pipe and bored in parallel or coaxially with the migration pipe.

[Brief explanation of drawings]

FIG. 1 is a functional explanatory diagram showing one embodiment of the isotachophoresis analyzer according to this invention. 1... Capillary type isotachophoresis analyzer, 2...
... Terminal electrolyte tank, 3 ... Sample injection port, 4 ... Potential gradient detector, 5 ... Electrolyte pushing/separating means, 6 ... ...Migration tubule, 7.
...Leading electrolyte tank, 8...Mountain motion control device.

Claims (1)

[Scope of utility model registration request] In an isokinetic electrophoresis analyzer equipped in this order with a terminal electrolyte tank, a migration conduit in which a sample injection port and a detector are installed, and a leading electrolyte tank, a distance A is provided from the detector to the leading electrolyte tank. A separation means is provided in the electrophoresis channel, and an electrolyte pushing means is provided in the migration channel or the leading electrolyte tank between the detector and the leading electrolyte tank, and based on the first detection of the target substance by the detector, The electrolyte pushing means is operated to push the target substance back from the detector to the sample injection port side by a distance B to continue electrophoresis, and based on the re-detection signal after a time T after receiving the first detection signal. , the time (A/
B) An isotachophoresis analyzer comprising an automatic control means for controlling the separation means to operate after T.