JPS6283643A - Sample injection apparatus - Google Patents

Abstract

PURPOSE:To make it possible to easily alter the injection amount of a specimen without losing the reproducibility of a column, by adjusting the amount of the specimen introduced into a capillary tube by changing the magnitude of voltage to be applied or a voltage applying time. CONSTITUTION:The magnitude of voltage to be applied or a voltage applying time is controlled by a controller E2' in such a state that the injection amount of a specimen from a sample valve S is made constant and the flow speed of the solvent supplied from a pump 7 is made constant to change the amount of the specimen introduced into a capillary tube 1. Thus injected specimen is separated in the tube 1 according to a theory of solubilization chromatography and a chromatograph corresponding to the solubilization ratio of the specimen is obtained by a detector 6. By this method, it is eliminated that the reproducibility of a column by injection work is lost, and the alteration of the injection amount of the specimen can be easily performed.

Description

[Detailed Description of the Invention] <Industrial Application Field> In recent years, a solvent consisting of a buffer solution and ionized micelles is flowed through a capillary tube, and the injected sample and the micelle dissolution phenomenon described in Ila, as well as the capillary electrophoresis method, have been studied. In addition, micellar solubilization chromatography has been proposed in which the sample is analyzed in one direction. The present invention relates to a sample 74 two-person apparatus suitable for such an analysis apparatus.

<Prior Art> FIG. 4 shows the original 1jJ4 configuration of solubilization chromatography. In the figure, numeral 1 indicates a thin tube constituting a column, for example, a fused silica capillary tube is used. 2 and 3 are containers containing a mixed solution of a buffer solution and micelles, and both ends of the kit Viralige 1 are inserted into these containers. For example, sodium dodecyl sulfate (S
Colloidal ions formed by dissolving DS) in the buffer solution are used.

E is a high voltage power supply, and the positive side is the electrode 4 immersed in the container 2.
In addition, the negative side is connected to the 11 pole 5 immersed in the container 3, and a high voltage is applied to both ends of the V-Village jet 1.

A detector 6, such as an ultraviolet spectrophotometer, is provided in a portion of the capillary duplex 1 near the negative pole 5.

With this (configuration), 4-11 Villa Dico-11 (
.: As shown in FIG. 55, two phases of micelles and M solution flow. That is, when a high voltage is applied, the buffer solution flows in the direction of the arrow through the electroosmotic moat. On the other hand, dissolved SDS (micelle) is an anion and tries to migrate to the positive side opposite to the flow of the buffer solution by electrophoresis, but the migration speed of the buffer solution is greater, so in the end, M
It reaches container 3 later than the buffer solution.

When a sample SM is injected from the positive electrode 4 side into the Kitobira Leech Cope 1 where such a two-phase flow exists, sample components that are not dissolved at all in the micelles ride the electroosmotic flow together with the buffer solution and flow rapidly. Move to the negative pole 5 side. On the other hand, the sample components completely dissolved and leaked into the micelles move at the same speed as the micelles, and τ-1ID tJpy <Container 3
Reach the side. In addition, the intermediate sample component that dissolves to some extent in the micelles moves at a moderate speed, and as a result, the sample component that moves the cavillary reach 1-1 depends on the rate of iD conversion. If this is detected by the detector 6, the chroma [-g is 1!] according to the solubilization rate of the sample. , t is received.

By the way, in such a case, there is a method of injecting sample 11 into the cavity pipe 1 by using the head (head FL), or by using the method of injecting the sample 11 into the cavity pipe 1. A method is used in which a high M power source is connected to the piping and the sample is injected using high voltage.

However, in the previous method, it is necessary to lift the sample container in order to obtain the desired I\d pressure, which makes the work complicated, and in the process of lifting the sample container, the capillary is often removed. I was trying to avoid deteriorating the reproducibility of the device by moving UP1. In addition, the rear right side had the disadvantage that it required an extra stage of high-voltage power supply for electrophoresis. In addition, none of these methods allowed precise control between sample injections and the ability to adjust sample injection 1:I) to the separation capacity of the column.

;Problems to be Solved by the Invention> The technical problems to be solved by the present invention are that in an analyzer using the capillary electrophoresis method, sample injection can be easily performed, and the column can be filled by 11 people. The object of the present invention is to realize a +1 sample injection device that does not impair reproducibility and allows the sample injection amount to be changed easily.

Means for Solving Problems〉 The structure of the present invention is based on hair! This is a number injection technique for an analyzer using II tube electrophoresis, the capillary -[
means for supplying a solvent such as a buffer solution from the flow side at a constant rate; means for depositing a fixed amount of sample into the piping connected to the capillary; and means for connecting the piping and the capillary and and a connection part for discharging the solution introduced into the capillary tube, and the sample injected into the pipe is sandwiched with the solvent and transferred to the connection part, and a voltage is applied to both ends of the capillary tube. ? :' The sample 1 was introduced into the 100 capillary tube by the flow formed by the S voltage! The suspension of the sample introduced into the capillary tube is adjusted by changing the length of the applied voltage or changing the application time of the applied voltage.

・Work> The above technical means works as follows. That is, when a sample is injected into the pipe, the sample is sandwiched by the solvent and forms a band, and is moved by the liquid supplied by the 7Fj medium supply means.

When the sample reaches the tip of the capillary, i! is present at both ends of the capillary. ′! If the i voltage is applied, a sample is introduced into the capillary tube by electroosmotic flow and electrophoresis.

When the amount of sample introduced into the pipe is constant and the flow rate of the solvent supplied from the solvent supply means is constant, the amount of the sample introduced depends on the magnitude of the applied voltage and the applied degree. It is determined by the application time of 1δ cat.

In the present invention, while the amount of the sample injected into the pipe is constant and the flow rate of the solvent supplied from the solvent supply means is constant, the thickness of the applied voltage is changed or the applied voltage is By changing the voltage application time, the sample fit introduced into the capillary tube is adjusted.

<Example> The present invention will be described in detail below with reference to the drawings. FIG. 1 is a block diagram showing an embodiment of the present invention, and FIG. 2 is a partially enlarged sectional view of the embodiment of the present invention shown in FIG.
Components that are substantially the same as those in FIG. 4 are given the same reference numerals, and explanations thereof will be omitted. Reference numeral 7 denotes a low-pressure tube that allows a mixed solution of the buffer solution and the mixture 11) to flow from the second container to the downstream side through piping 8, and constitutes a solvent supply means.

The part E' surrounded by the dashed line is a high voltage power supply section, which applies high voltage from the high voltage power supply "1' to both ends of the capillary tube 1 via the 1iIJ III device E2'. S is the piping 8 A sample valve provided in the middle of the pipe (composed of a switching valve s1 and a measuring loop S2) injects a fixed amount of the sample supplied from the sample injection port S3 or 13 into the pipe 8 by switching the valve s1. .

A tube 9 is connected to the end of the pipe 8 (connected to the positive side of the high voltage 7If source E').

Reference numeral 10 denotes an L-type connecting portion that connects the pipe 8 and the capillary duplex 1 and discharges the solution that has not been introduced into the capillary tube 1.

Figure 2 shows an enlarged cross section of this part. In the figure, 10a is T
The main body of the type connection part, 10b is a tightening screw that is screwed into the -1ii1 opening and fixes the tubular electrode 9 to the main body 10a.
10c is a square opening [screwed into part 1 and gear pillary tube 1]
A tightening screw 10d fixes the jJl liquid duplex 11 to the main body 10a. Note that the capillary duplex 1 is fixed with its tip inserted into the duplex-shaped Ti pole 9.

In this way, regarding the operation of 'A iF? made by M,
The explanation will be given according to the explanatory diagram in FIG. In this figure, the first
Elements that are the same as those in J3 in the figure are given the same reference numerals. FIG. 3(a) shows the state before the sample is injected into the pipe 8. In this state, only the buffer solution, the micelles, and the solvent are flowing through the pipe 8 at a constant speed, and the solvent that has not been introduced into the capillary tube 1 is discharged from the drain tube 11.

When a certain amount of sample is injected into the power distribution 8 from the rimpull valve S, the sample is made into a lean switch (band B) by the sea urchin solvent shown in FIG. It moves at a constant speed.

When the band B reaches the tip of the capillary duplex 1, if a high voltage is applied to both ends of the capillary duplex 1, the sample is introduced into the capillary tube 1 by electroosmotic flow and electrophoresis.

When the sample nff1 injected into the piping 8 through the rimple valve S is constant and the flow rate of the solvent supplied from the pump 7 is constant, the suspension of the introduced sample is determined by the magnitude of the applied voltage,
It also depends on the application time of the applied voltage. In the present invention, the sample injection and output from the sample valve S is kept constant,
While the flow rate of the solvent supplied from the pump 7 is kept constant, the magnitude or application time of the applied voltage is controlled by the controller E2' to change the amount of sample introduced into the chitopillary tube 1. There is.

The sample injected in this way is separated in the capillary tube 1 according to the principle of solubilization chromatography as explained above, and the detector 6 obtains a chromatogram according to the solubilization rate of the sample. There is.

<Effects of the Invention> According to the present invention, since the sample can be automatically injected into the capillary duplex, the work is simplified, and J.
The reproducibility of the device was increased by +11 due to the fact that the IlOl pillary tube was moved during the rippling injection process.
It never happens. Furthermore, since the optimal sample injection layer can be selected according to the separation capacity of the column, analysis can be performed without reducing column efficiency.

In addition, in the explanation of the present invention so far, the case where the present invention is implemented on a micelle solubilized Chroma 1 Hegelafy has been explained, but the present invention is not limited to this, and can also be applied to an analysis device based on other capillary aerophoresis methods. The invention can be practiced with equal ease.

[Brief explanation of drawings]

FIG. 1 is a block diagram showing an embodiment of the present invention, FIG. 2 is a partially enlarged sectional view of the embodiment of the present invention shown in FIG. 1, and FIG. 3 is an embodiment of the present invention shown in FIG. Explanatory diagram of the operation of the device, No. 4
The figure is an actual diagram of the solubilization chromatography, and FIG. 5 is an explanatory diagram of the operation of the solubilization chromatography shown in FIG. DESCRIPTION OF SYMBOLS 1... Capillary tube, 2, 3... Solvent container, 6... Detector, 7... Pump, 8... Piping, 9... Tubular electrode, 10... F type connection part , 11...Drainage duplex, E'...High voltage power supply section, El'...High voltage?
W source, E2'...controller, S...sample valve Fig. 3

Claims (1)

[Claims] A sample injection device for an analytical device using capillary electrophoresis, which includes means for supplying a solvent such as a buffer solution at a constant rate from the upstream side of the capillary tube, and a fixed amount of solvent such as a buffer solution to a pipe connected to the capillary tube. and a connection part that connects the piping and the capillary tube and discharges a solution that has not been introduced into the capillary tube, and a state in which the sample injected into the piping is sandwiched with the solvent. In an apparatus in which the sample is transferred to the connecting part and introduced into the capillary tube by a flow formed by a high voltage applied to both ends of the capillary tube, the magnitude of the applied voltage is A sample injection device characterized in that the amount of sample introduced into the capillary tube is changed by changing the applied voltage or by changing the application time of the applied voltage.