JPS6333662B2 - - Google Patents

Description

[Detailed Description of the Invention] The present invention provides an electrophoresis analyzer, in particular, an electrophoresis analyzer capable of cutting unnecessary polymer components (components with large particle diameters) from among the components contained in a sample for electrophoresis. This invention relates to an electrophoresis analyzer.

Electrophoresis that connects the liquid phases of a liquid containing charged particles with high mobility (hereinafter abbreviated as leading liquid) and a liquid containing charged particles with low mobility (hereinafter abbreviated as terminal liquid). In an electrophoresis analyzer that is configured to inject a sample into the boundary between both liquids in the tube and apply a high voltage to both ends of the migration tube, all ionic components with mobility are separated by their own mobility. Separated and detected. However, in order to achieve clear separation, the mobility of each component must be between the mobility of charged particles in the leading liquid and the mobility of charged particles in the terminal liquid, and the mobility of each component must be about 2% between them. The above difference is a necessary condition.

By the way, serum components, for example, contain a large number of components, ranging from high-molecular substances such as proteins to low-molecular substances such as organic acids and inorganic ions. It is difficult to make the difference in mobility between components 2% or more. Therefore, conventional electrophoretic analyzers tend to provide insufficient separation.

Therefore, if the purpose is only to separate and detect low-molecular substances, and if the high-molecular substances can be cut and subjected to electrophoresis, it is necessary to select the pH of the electrolyte that is appropriate for the mobility of each component of the low-molecular substances. As a result, it is possible to obtain a value between the mobility of the leading liquid charged particles and the mobility of the terminal liquid charged particles, which has a difference of 2% or more from each other, so that each component can be clearly separated. will be possible. Moreover, if the polymeric substances are removed, contamination of the electrophoresis tube and detector is prevented, and a decrease in separation performance is prevented, so that good separation can be achieved in this sense as well.

If the macromolecular components in the sample can be cut out and subjected to electrophoresis in this way, the analysis results can be positively influenced. This type of electrophoresis analysis method is not only suitable for analyzing low-molecular substances in serum samples, but also for analyzing the molecular weight of samples, such as when analyzing polymers sampled during the polymerization process. It can be applied very effectively even to components of a sample that contain many ions with different values.

The purpose of the present invention is to provide an apparatus suitable for electrophoretic analysis as described above. The present invention is characterized in that a filter mounting mechanism is provided in which more than one filter member can be selectively interposed, and unnecessary polymer components in a sample can be cut out by the filter members for electrophoresis.

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

In FIG. 1, reference numeral 1 denotes an electrophoresis analyzer according to an embodiment of the present invention, which includes a high voltage power supply 2, a terminal liquid electrode tank 3, a sample introduction section 4, a migration tube 5,
and a filter mounting mechanism 8 that can selectively interpose filters 12a, 12b, 12c, and 12d on the detector 6, the leading liquid electrode tank 7, and the migration tube 5 between the sample injection section 4 and the detector 6. It is configured. S is a switch inserted in the high voltage circuit, and M is a syringe that supplies the sample to the sample introduction section 4.

The filter mounting mechanism 8 is as shown in FIG.
It is composed of two fixed terminals 9 and 11 that are integrally connected to the migration tube 5, and a disk-shaped rotor 10 that is rotatably and slidably clamped between them. The terminals 9 and 11 have through holes 9a and 11, respectively, at eccentric positions connected to the migration tube 5.
a is output. On the other hand, the rotor 10 has a through hole 9 from the rotation center O as shown in FIG.
The required number of filter mounting holes 10, for example, 4, are formed on the circumference of radius r reaching the center O' of a and 11a.
a, 10b, 10c, and 10d are provided.
Each of these filter mounting holes 10a, 10b,
10c and 10d are replaceable with gel-like filters 12a, 12b, 12c, and 12d, such as agarose, which selectively separate components in the sample according to particle size, in other words, molecular weight. is filled with. Reference numeral 13 denotes connectors that connect the migration tube 5 and the fixed terminals 9 and 11, respectively.

In FIG. 2, the filter attachment hole 10a of the rotor 10 is inserted into the migration tube 5, but the rotor 10 is rotated around the rotation center O.
By rotating 90 degrees, you will see filter mounting hole 10.
b, 10c, and 10d are successively installed in the electrophoresis tube 5. Therefore, the filters 12a, 12
If the molecular weights that can be separated by b, 12c, and 12d are sequentially increased, electrophoresis can be performed by cutting out components having a desired size or more by switching adjacent filters. In addition,
Since the charged particles contained in the leading liquid and the terminal liquid are low molecular ions, it goes without saying that they can freely pass through each of the filters 12a, 12b, 12c, and 12d.

Next, the operation of this embodiment apparatus 1, that is, the analysis operation will be explained.

In FIG. 1, a sample is injected into the boundary between the leading liquid and the terminal liquid through a rubber septum (not shown) of the sample introduction part 4 using a syringe M, and then the switch S is closed and electrophoresis is performed. Among the contained ion components, only low-molecular ions of a size that can pass through the filter 12a are migrated to the detector 6 and analyzed, but migration of higher molecular ions is blocked.

Figure 4 shows a separation electrophoretogram when the sample components are not cut with a filter, and Figure 5 shows a separation electropherogram when the sample is cut with a filter, and the separation ability is significantly improved. I understand that. That is, FIG. 5 is part A of FIG. 4, and part B has been cut out.

In the above embodiment, a case has been described in which the filter mounting mechanism is provided separately from the sample introduction section, but it goes without saying that a structure in which the two are combined together may also be used.

As is clear from the above explanation, according to the present invention, it is possible to perform electrophoresis of the sample by cutting out unnecessary polymeric components in the sample, thereby preventing contamination of the electrophoresis tube and detector by the polymeric components. It is possible to perform analysis with good resolution.

[Brief explanation of the drawing]

Fig. 1 is an explanatory diagram of the configuration of one embodiment of the electrophoretic analyzer of the present invention, Fig. 2 is a perspective view of the filter mounting mechanism, Fig. 3 is a front view of the rotor of the filter mounting mechanism, and Fig. 4 is a sample Fig. 5 is a separation electrophoretogram when the sample is not separated by a filter. 1... Electrophoresis analyzer, 4... Sample introduction section,
5... Electrophoresis tube, 6... Detector, 8... Filter mounting mechanism, 12a, 12b, 12c, 12d...
filter.

Claims (1)

[Claims] 1. A sample is injected into the boundary between the two liquids in an electrophoresis tube that connects the liquid phases of a liquid containing charged particles with high mobility and a liquid containing charged particles with low mobility. In an electrophoresis analyzer configured to apply a high voltage to both ends of an electrophoresis tube, a filter attachment mechanism is provided in the electrophoresis tube from the sample introduction part to the detector, and the filter member attachment mechanism is attached to the electrophoresis tube. A through hole is provided in the middle and forms part of the upper and downstream sides of the migration channel, respectively.
Two fixed terminals facing each other with a space between them, and a plurality of fixed terminals that are rotatably and slidably sandwiched between these fixed terminals and spaced apart from each other on the circumference around the center of rotation. It has a filter mounting hole, and one of the filter mounting holes, which can be switched and selected by rotation, allows communication between the two through holes, and the remaining filter mounting hole is used as a sliding surface with both fixed terminals. An electrophoretic analyzer comprising: a rotor that can be sealed tightly; and filters that are replaceably filled in each of the filter mounting holes and that allow separation of different molecular weights.