JPH08201362A - Automatic pre-column deriving device - Google Patents

Abstract

PURPOSE: To provide an automatic pre-column deriving device capable of satisfactorily deriving samples. CONSTITUTION: This automatic pre-column deriving device is provided with multiple racks 21-23 capable of holding containers, an automatic moving mechanism 4 capable of moving a needle sucking and discharging a liquid, and a temperature regulating mechanism capable of independently regulating the temperatures of at least two of the racks 21-23. The automatic moving mechanism 4 moves samples and a deriving reagent between the racks 21-23 individually temperature-set by the temperature regulating mechanism and the containers to derive the samples.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to an automatic precolumn derivatization apparatus for use in high performance liquid chromatography.

[0002]

2. Description of the Related Art In a method for separating substances by utilizing a difference in affinity between a solid phase and a mobile phase, liquid chromatography is one in which a liquid is used as a mobile phase. High performance liquid chromatography is a method in which the mobile phase is accelerated by a pump having a large discharge pressure in this liquid chromatography. In such liquid chromatography, derivatization by reacting with a reagent is carried out for the purpose of enhancing the selectivity for the desired reactivity of the detector and the detection sensitivity. Pre-column derivatization is a derivatization that is injected into the column and preceded by chromatographic separation. Generally, in high performance liquid chromatography, various analyzes are performed on a large number of samples, and therefore, automation of the precolumn derivatization process is desired. Further, this derivatization reaction is a chemical reaction carried out between the sample and the derivatization reagent, and it is desirable to set the reaction temperature suitable for derivatization.

Conventionally, as an automatic pre-column derivatization device which meets such requirements, for example, a device using a tube as shown in FIG. 6 is known. This conventional automatic pre-column derivatization apparatus sucks the sample and the derivatization reagent into a tube before mixing into the tube, mixes them both in the tube, and heats them by a heating means such as a micro oven to control the reaction rate. However, the reaction conditions are adjusted.

[0004]

However, the conventional automatic precolumn derivatization device has a problem that it is difficult to perform good derivatization. For example, the sample and the derivatization reagent are simply mixed by diffusion in the tube, so that the mixing tends to be insufficient, resulting in a problem in reproducibility of analysis. Moreover, since the mixing and the heating reaction are performed in the same place,
During the heating reaction, there is a drawback that other samples cannot be mixed and the analysis efficiency is lowered.
Therefore, it is an object of the present invention to provide an automatic pre-column derivatization apparatus that can solve the above-mentioned conventional problems and perform good derivatization.

[0005]

According to the present invention, a plurality of racks capable of holding a container, an automatic moving mechanism capable of moving a needle capable of sucking and discharging a liquid, and at least two of the racks are provided. Equipped with a temperature control mechanism that can independently control the temperature of two racks.The automatic movement mechanism moves the sample and derivatization reagent between racks and containers that are individually temperature-controlled by the temperature control mechanism. The above object is achieved by performing derivatization of the sample. The automatic pre-column derivatization apparatus of the present invention is an apparatus for automatically derivatizing a sample with a derivatization reagent before injecting an analytical sample into a column. The sample used for derivatization, the derivatization reagent, and the derivatization reagent This is an apparatus in which a rack for storing reacted samples and the like is provided in the apparatus. The automatic transfer mechanism of the present invention is a mechanism for automatically transferring a sample or derivatization reagent between containers according to a preset procedure and time, thereby performing mixing or derivatization reaction in the container. The temperature control mechanism of the invention controls the temperature of each of the racks for the containers containing the sample, the derivatization reagent, and the sample subjected to the derivatization reaction independently of each other, thereby allowing the sample, the derivatization reagent, and the derivatization reaction to be performed. It is a mechanism that makes it possible to adjust the temperature of the sample and the like.

In the first embodiment of the present invention, a container containing a sample, a container containing a derivatization reagent, an empty container and a container containing a sample during the derivatization reaction are held in individual racks, and a temperature control mechanism is provided. Allows the temperature of all or at least two of these racks to be independently adjusted, whereby the temperature of the sample, the derivatization reagent, and the sample during the derivatization reaction can be individually controlled. In the second embodiment of the present invention, a container containing a sample, a container containing a derivatization reagent, an empty container, and a container containing a solution in the derivatization reaction are held in individual racks, and the temperature control mechanism is All of these racks or at least two temperatures can be adjusted independently,
This allows individual temperature control of the sample, derivatization reagent, solution during the derivatization reaction, and
Mixing and derivatization reactions can be performed independently.

In a third embodiment of the present invention, the automatic moving mechanism is provided with a mechanism for moving a container, whereby the container after mixing can be moved to a rack for carrying out a derivatization reaction. it can. In the fourth embodiment of the present invention, the container moving mechanism of the automatic moving mechanism uses a magnetic force, and moves the container by, for example, a magnetic attraction force between a magnetic body and an electromagnet. As a result, the automatic movement mechanism can be used for both movement of the liquid and movement of the container.

[0008]

In the automatic precolumn derivatization apparatus of the present invention, before the precolumn derivatization, each rack in the apparatus is provided with a container in which a sample is injected, a container in which a derivatization reagent is injected, and an empty container. Store it. The temperature of each of these racks is independently adjusted by a temperature adjusting mechanism to suppress changes in the sample and derivatization reagent in the container. The automatic moving mechanism extracts the sample and the derivatization reagent from each container, moves them into another container, and mixes in the container. After the derivatization reaction of the mixed solution, it is injected into a column for chromatographic analysis. During this derivatization reaction, the temperature adjusting mechanism of the present invention can adjust the temperature of the container for derivatization to a temperature suitable for derivatization. At this time, the pre-reaction sample and derivatization reagent held in the same automatic pre-column derivatization apparatus are adjusted to a temperature different from this derivatization, and are not affected by the temperature change due to derivatization. .

When the automatic pre-column derivatization apparatus of the present invention is provided with a rack for empty containers and a rack for containers for storing the solution in the derivatization reaction, the automatic transfer mechanism is an empty container. The sample can be mixed with the derivatization reagent in the container, and the mixed solution can be derivatized in the container for the derivatization reaction. In the case of this configuration, mixing and derivatization can be adjusted to different temperatures, and the time for derivatization can be set accurately.

Further, when the automatic moving mechanism has a mechanism for moving the container, the moving process for moving the mixed container to the rack for derivatization reaction is performed by the automatic moving mechanism for moving the liquid. Therefore, the movement of the liquid and the movement of the container can be combined by one automatic movement mechanism.

[0011]

[First Embodiment of the Invention]

(Configuration of First Embodiment) FIG. 1 is a schematic configuration diagram for explaining a configuration of a first embodiment of the present invention. In FIG.
The automatic pre-column derivatization apparatus 1 includes a rack section 2 having a plurality of racks, a base 3 supporting the rack section 2,
The moving mechanism 4 is provided for moving the needle 41 between the rack portion 2 and the container. In FIG. 1, the rack unit 2 includes a first rack 21, a second rack 22, and a third rack 23. These racks are installed on the base 3, and the temperature control mechanism built in the base 3 enables independent temperature control. The racks 21 to 23 have a mechanism for storing containers,
The type of container to be stored can be set arbitrarily. In this embodiment, for example, the first rack 21 holds the reagent container 25 in which the derivatization reagent is injected, the second rack 22 holds the sample container 26 in which the sample is injected, and the third rack The rack 23 holds a reaction container 27 in which an empty or reacting solution is injected.

Therefore, in the first embodiment, the first
The rack 21 is used as a reagent rack, and the second rack 22
Is used as a sample rack, and the third rack 23 is used as a reaction rack. Further, each container is formed so as to be able to be put in and taken out from a storage section formed in each rack, and further has a structure into which the needle 41 of the moving mechanism 4 can be inserted.

The temperature adjusting mechanism can independently adjust the temperature of at least two of the racks, and each rack is provided with a temperature display section 31 and a temperature setting section 32. The temperature display section 31 can use any other display means such as a liquid crystal display means, and the temperature setting section 32 can use any other setting means such as a temperature increase button and a temperature decrease button. The temperature of each rack can be set by operating the temperature setting unit 32, and can be confirmed by the set temperature displayed on the temperature display unit 31. Further, the temperature display unit 31 can also display the actual temperature of the rack by mode conversion. In this first embodiment, the first rack 2
An example in which the temperature of all the racks 1 to 3 is adjustable is shown. Further, as the temperature adjusting mechanism, for example, a Peltier element is used as the temperature changing means,
It can be formed by a structure in which the surface of the rack is surrounded by an aluminum block. When this Peltier element is used, the temperature can be adjusted, for example, from 4 ° C to 70 ° C.

Further, the moving mechanism 4 is provided with a needle 41 capable of sucking and discharging the liquid so that the liquid can be taken in and out of the container, and the needle 41 is XY.
It can be moved three-dimensionally by the Z drive unit 42.
The XYZ drive unit 42 can use a normal XYZ drive mechanism. The needle 41 driven by the moving mechanism 4 moves between the racks to move the sample and the reagent in the container, and performs mixing and derivatization processing. In addition, the solution derivatized by the automatic precolumn derivatization apparatus of the first embodiment is injected into the column through the injection port 51 of the column injection unit 5, and the chromatographic analysis is performed. The order of movement of the moving mechanism 4 and the temperature adjustment of the temperature adjusting mechanism can be set in advance according to the reaction content, and the automatic precolumn derivatization apparatus can automatically perform processing according to the setting. .

(Operation of First Embodiment) Next, the operation of the first embodiment of the present invention will be described with reference to FIG. FIG. 2 is a diagram for explaining the operation procedure of the first embodiment. Figure 2
For example, pre-column derivatization in the case where the sample is lauric acid and the derivatization reagent is 9-anthryl diazo metal (ADAM reagent) is shown. Note that FIG. 2 shows the procedure in alphabetical order (lowercase).
In FIG. 2, it is assumed that the reagent container 25 is stored in the first rack 21, the sample container 26 is stored in the second rack 22, and the reaction container 27 is stored in the third rack 23. . The temperature setting unit 32 is operated to set the temperature of each rack while checking the set temperature displayed on the temperature display unit 31. When the actual temperature of the rack is displayed after the temperature setting is completed, mode conversion is performed and the actual temperature of the rack is displayed on the temperature display unit 31. Further, the moving procedure of the moving mechanism is set using a setting device (not shown). This setting includes, for example, setting of a sample to be derivatized and its reagent, specification of a reaction container for injecting a solution to be mixed, derivatization time, and the like.

First, in FIG. 2A, the second rack 22 containing the sample is set to, for example, 4 ° C. by the temperature adjusting mechanism, and the third rack 23 containing the empty container is set.
Is set to 40 ° C., for example. Then, the XYZ drive unit 42 is driven to move the needle 41 to the second rack side,
The sample is inserted into the sample container 26 to suck up the sample (a in the figure). Next, while holding the sucked up sample, it is pulled up from the sample container 26 (b in the figure) and moved to the third rack 23 side (c in the figure). The needle 41 is lowered to the set reaction container 27 on the side of the third rack 23 (d in the figure), and the sample is discharged into the reaction container 27. As a result, the sample is injected into the reaction container 27. After the injection of this sample, the needle 41 is washed (not shown).

Next, in FIG. 2B, the XYZ drive unit 42 is driven to move the needle 41 to the side of the first rack 21 in which the reagent is stored (e in the figure), and is placed in the reagent container 25. Insert and suck up the reagent (f in the figure). Next, while holding the sucked reagent, the reagent container 25 is pulled up (g in the figure) and moved to the third rack 23 side (h in the figure).

A reaction container 27 in which a sample is injected.
On the other hand, the needle 41 is lowered (i in the figure) to discharge the reagent into the reaction container 27. As a result, the sample and the reagent are injected into the reaction container 27. After the injection of this reagent, the needle 41 is washed (not shown).

Next, in FIG. 2C, the XYZ drive unit 42 is driven to move the needle 41 again.
7, and suction and discharge of the sample and reagent in the reaction container 27 are repeated to perform mixing (j in the figure). The number of repetitions of suction and discharge can be set in advance, and can be set to 3 times, for example. After mixing the sample and the reagent by this mixing, 1
The derivatization reaction is allowed to occur for about 0 minutes.

Next, referring to FIG. 2D, the XYZ driving section 42 is driven to move the needle 41 again.
7, and the derivatized solution is sucked from the reaction container 27. While holding the sucked solution, the XYZ driving section 42 is driven to pull up the needle 41 to move it to the column injection section 5 side (k in the figure), and the solution is added to the column injection section 5
Inject (l in the figure). This completes the precolumn derivatization of one sample. Thereafter, the precolumn derivatization of another sample is performed by the same steps as described above.

(Effects of the First Embodiment) Since the temperature can be adjusted independently for each rack by the temperature adjusting mechanism, the sample and the reagent do not change due to the temperature required for the derivatization reaction and are in a stable state. Can be maintained, and reproducibility and analysis accuracy can be improved even in long-term analysis.

Second Embodiment of the Present Invention (Configuration of Second Embodiment) FIG. 3 is a schematic configuration diagram for explaining the configuration of the second embodiment of the present invention. In FIG.
The automatic pre-column derivatization apparatus 1 of the second embodiment has a rack section 2 including a plurality of racks, as in the first embodiment.
And a moving mechanism 4 for moving the needle 41 between the rack portion 2 and the container. In FIG. 3, the rack unit 2 of the second embodiment includes a first rack 21, a second rack 22, a third rack 23, and a fourth rack 24. These racks are installed on the base 3, and the temperature control mechanism built in the base 3 enables independent temperature control. The racks 21 to 24 have a mechanism for storing the containers, and the type of the containers to be stored can be set arbitrarily. In the second embodiment, for example, the reagent container 25 in which the derivatization reagent is injected into the first rack 21.
The second rack 22 holds the sample container 26 in which the sample is injected, the third rack 23 holds the empty container 26, and the fourth rack 24 is filled with the solution during the reaction. The reaction container 29 that is being held is held.

Therefore, in the second embodiment, the first
The rack 21 is used as a reagent rack, and the second rack 22
Is used as a sample rack, the third rack 23 is used as an empty container rack, and the fourth rack 24 is used as a reaction rack. Further, each container is formed so as to be able to be put into and taken out from the storage portion formed in each rack, similarly to the first embodiment, and further has a structure into which the needle 41 of the moving mechanism 4 can be inserted.

The temperature adjusting mechanism may have the same structure as that of the first embodiment, and at least two of the racks can be independently temperature adjusted. Second
In the embodiment, an example is shown in which the temperature of all the racks from the first rack 21 to the third rack 24 can be adjusted. In addition,
The temperature adjusting function can be omitted for the third rack 23 that stores empty containers. As the temperature adjusting mechanism, a Peltier element may be used as in the first embodiment, and the surface of the rack may be surrounded by an aluminum block.

The moving mechanism 4 of the second embodiment can be provided with a mechanism for moving the container. FIG. 4 is a view for explaining the container moving mechanism of the second embodiment. The container moving mechanism 41 can be constituted by, for example, an electromagnet provided in the vicinity of the needle 41, and the needle 41 of the container 6
It can be configured to cooperate with the container moving mechanism 42 such as a magnetic body provided in a portion opposed to. The moving mechanism 4, the order of movement of the moving mechanism 4, and the temperature adjustment of the temperature adjusting mechanism can be performed in the same manner as in the first embodiment.

(Operation of Second Embodiment) Next, the operation of the second embodiment of the present invention will be described with reference to FIG. FIG. 5 is a diagram for explaining the operation procedure of the second embodiment. Similarly to FIG. 2, FIG. 5 also shows pre-column derivatization when the sample is lauric acid and the derivatization reagent is 9-anthryldiazometal (ADAM reagent). Note that FIG. 5 shows the procedure in alphabetical order (lowercase). In FIG. 5, the reagent container 25 is stored in the first rack 21, the sample container 26 is stored in the second rack 22, the empty container 28 is stored in the third rack 23, and the fourth rack 24 is stored. It is assumed that the reaction container 29 is stored. Then, similarly to the first embodiment, the temperature setting and the moving procedure of the moving mechanism are set.

First, in FIG. 5A, the second rack 22 containing the sample is set to, for example, 4 ° C. by the temperature adjusting mechanism, and the fourth rack 24 containing the reaction container 29 is set to, for example, 40 ° C. Keep it. The empty container 28
The temperature of the third rack 23 in which is stored is kept at room temperature or set to 4 ° C. like the second rack 22. Then, the XYZ drive unit 42 is driven to move the needle 41 to the second rack side, and is inserted into the sample container 26 to suck up the sample (a in the figure). Next, while holding the sucked up sample, it is pulled up from the sample container 26 (b in the figure) and moved to the third rack 23 side (c in the figure). Third
The needle 41 is lowered to the set empty container 28 on the rack 23 side (d in the figure), and the sample is discharged into the empty container 28. As a result, the sample is injected into the reaction container 27. After the injection of this sample, the needle 41 is washed (not shown).

Next, in FIG. 5B, the XYZ drive section 42 is driven to move the needle 41 to the side of the first rack 21 in which the reagent is stored (e in the figure), and is placed in the reagent container 25. Insert and suck up the reagent (f in the figure). Next, while holding the sucked reagent, the reagent container 25 is pulled up (g in the figure) and moved to the third rack 23 side (h in the figure). Needle 4 for empty container 28 in which the sample is injected
1 is lowered (i in the figure), and the reagent is discharged into the empty container 28. As a result, the sample and the reagent are injected into the empty container 28. After the injection of this reagent, the needle 41 is washed (not shown).

Next, in FIG. 5C, the XYZ driving section 42 is driven to insert the needle 41 into the empty container 28 in which the sample and the reagent are injected again, and the sample and the reagent in the empty container 28 are inserted. Mixing is performed by repeating suction and discharge of and (j in the figure). The number of repetitions of suction and discharge can be set in advance, and can be set to 3 times, for example.

Next, in FIG. 5D, the XYZ driving section 42 is driven to move the first container moving mechanism 43 to the empty container 28 side (k in the figure), and the second container moving mechanism of the container. 61 approaches or abuts. At this time, the container can be moved by the moving mechanism 4 by the cooperation of the first container moving mechanism 43 and the second container moving mechanism 61. FIG. 4B
Indicates the one for cooperative operation between the first container moving mechanism 43 and the second container moving mechanism 61, and is indicated by the arrow b after the first container moving mechanism 43 and the second container moving mechanism 61 are joined by the arrow a. As described above, the container can be moved. When the first container moving mechanism 43 is an electromagnet, the containers can be joined by supplying an exciting current to the electromagnet. Further, when the container is separated from the moving mechanism, the supply of the exciting current to the electromagnet may be stopped.

The moving mechanism 4 moves the empty container 28 in which the sample and the reagent are injected to the side of the fourth rack 24 (l,
m), the empty container 28 is lowered to the storage place in the fourth rack 24 (n in the figure). Then, the container moving mechanism is opened and the empty container 28 is placed in the fourth rack 24. After that,
This empty container 28 is referred to as a reaction container 29.

Next, in FIG. 5E, the reaction container 2
The samples and reagents in 9 start the derivatization reaction in the fourth rack 24 where the temperature for derivatization is set. Since the temperature of the third rack is unsuitable for the derivatization reaction, the derivatization reaction starts when the reaction container 29 moves into the fourth rack 24. Then, the reaction container 29 is placed in the fourth rack 24 for a preset reaction time.

Next, in FIG. 5F, the XYZ drive unit 42 is driven to insert the needle 41 into the reaction container 29 (o in the figure), and the derivatized solution is sucked from the reaction container 29. To do. Then, while holding the aspirated solution,
The XYZ drive unit 42 is driven to pull up the needle 41 (p in the drawing) and moved to the column injection unit 5 side (q in the drawing).
The solution is injected into the column injection part 5 (r in the figure). After the injection, the reaction container 29 is returned to the third rack. This completes the precolumn derivatization of one sample. Thereafter, the precolumn derivatization of another sample is performed by the same steps as described above.

(Effects of the Second Embodiment) As in the first embodiment, since the temperature can be adjusted independently for each rack by the temperature adjusting mechanism, the sample or reagent can be adjusted depending on the temperature required for the derivatization reaction. Does not change and a stable state can be maintained, and reproducibility and analysis accuracy can be improved even in long-term analysis. In addition, since the derivatization reaction of the solution after mixing can be performed independently from the mixing, the reaction time of derivatization can be set accurately, and the reproducibility and accuracy of analysis can be improved. it can. Moreover, since the temperature of the fourth rack for carrying out the derivatization reaction can be set for each reaction by the temperature control mechanism, the derivatization for each reagent can be performed more accurately.

(Modification) As the container moving mechanism, a mechanism using air suction force instead of magnetic force can be used. In this case, for example, an intake port is provided on the moving mechanism side, The container can be moved by suctioning the edges.

[0036]

As described above, according to the present invention,
It is possible to provide an automatic precolumn derivatization apparatus that can perform good derivatization.

[Brief description of drawings]

FIG. 1 is a schematic configuration diagram for explaining a configuration of a first embodiment of the present invention.

FIG. 2 is a diagram for explaining the procedure of the operation of the first embodiment of the present invention.

FIG. 3 is a schematic configuration diagram for explaining a configuration of a second embodiment of the present invention.

FIG. 4 is a view for explaining the container moving mechanism of the second embodiment of the present invention.

FIG. 5 is a view for explaining the procedure of the operation of the second embodiment of the present invention.

FIG. 6 is a schematic diagram of a conventional automatic precolumn derivatization apparatus.

[Explanation of symbols]

1 ... Automatic pre-column derivatization device, 2 ... Rack unit, 4 ...
Moving mechanism 21, 22, 23, 24 ... Rack, 25 ... Reagent container, 26 ... Sample container, 27 ... Reaction container, 28
... reaction vessel.

Claims (1)

[Claims] 1. A plurality of racks capable of holding containers, an automatic moving mechanism capable of moving a needle capable of sucking and discharging a liquid, and temperature control of at least two racks in the racks independently. And a temperature adjusting mechanism capable of performing derivatization of the sample by moving the liquid in the container between the rack and the container whose temperature is individually set by the temperature adjusting mechanism. An automatic precolumn derivatization device characterized by the above.