JP2010169613A - Method and assist device for analyzing liquid composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for analyzing liquid compositions using a solid phase extracting column, conditioning the solid phase extracting column with a small amount of solvent, and replacing air existing around solid phase adsorbed particles with the solvent sophisticatedly with few variations, and also to provide an assist device for analyzing the liquid components, simply performing the above operation. <P>SOLUTION: The method for analyzing the liquid components uses the solid phase extracting column 3 having a packed bed 34 filled with a solid phase adsorbent for adsorbing components contained in a liquid sample, has at least a conditioning step of supplying a replaceable solvent devoid of sample components to wet the packed bed 34 with the replaceable solvent, and in the conditioning step, promotes elimination of air contained in the packed bed 34 by placing the entirety of the solid phase extracting column 3 in a reduced pressure environment. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a liquid component analysis method and a liquid component analysis auxiliary device, and more particularly, to a liquid component analysis method using a solid phase extraction column and a liquid component analysis using a solid phase extraction column. The present invention relates to a liquid component analysis auxiliary device.

  As a pretreatment for concentrating or purifying components to be analyzed contained in a liquid sample, a treatment using a solid phase extraction column is widely performed (see, for example, Patent Document 1). The column for solid phase extraction is one in which a porous container made of polypropylene or the like is filled with porous particles such as silica or polymer as a solid phase adsorbent. The solid phase extraction column includes a packed bed filled with a solid phase adsorbent, a liquid holding unit for holding the liquid above the packed bed, and an outlet for discharging the liquid below the packed bed. The liquid supplied to the liquid holding unit passes through the packed bed of the solid phase extraction column and flows out from the outlet.

  In order to concentrate and purify the target component using the solid phase extraction column, first, the sample solution is supplied to the liquid holding part of the solid phase extraction column, and the sample solution is packed into the packed bed of the solid phase extraction column. The target component in the sample solution is adsorbed to the solid phase adsorbent by passing the sample. Thereafter, the target component adsorbed by the solid phase adsorbent inside the packed bed is eluted by flowing the eluate through the column. The eluted target component is then subjected to various analyzes such as liquid chromatography.

  By the way, the solid phase adsorbent of the solid phase extraction column is dry-filled. For this reason, before the sample solution is adsorbed on the solid phase adsorbent, an operation of wetting the packed bed containing the solid phase adsorbent using an appropriate solvent is performed. This operation is called conditioning, and by appropriately performing conditioning, the air present around the solid-phase adsorbent particles is replaced with the solvent, and the surface of the solid-phase adsorbent particles is wetted with the solvent. As a result, the sample solution containing the target component can sufficiently come into contact with the surface of the solid-phase adsorbent particles, and the solid-phase adsorbent exhibits sufficient adsorption performance. On the other hand, if the conditioning is insufficient, air remains around the solid-phase adsorbent particles, and the adsorption ability of the solid-phase adsorbent is not sufficiently exhibited.

  Conditioning is generally performed by reducing the pressure on the outlet side while supplying a solvent to the liquid holding part of the solid phase extraction column. Thus, by providing a pressure difference between the liquid supply side and the discharge port side of the column, the solvent quickly passes through the packed bed of the solid phase extraction column, so that the time required for conditioning can be shortened.

JP-A-6-258203

  However, it is difficult to completely replace the air present around the solid-phase adsorbent particles existing inside the packed bed with a solvent, and there is a problem that a large amount of solvent and time are required to completely replace the air. Exists. On the other hand, if the air around the solid-phase adsorbent particles is not completely replaced by the solvent, the adsorption performance of the solid-phase adsorbent will not be fully demonstrated, and the expected analysis results will be obtained. There may be no. Furthermore, since it is impossible to perform conditioning while observing the amount of air present around the solid-phase adsorbent particles, the degree of substitution of the air present around the solid-phase adsorbent particles with the solvent is suitable for solid-phase extraction. Often it varies from column to column. For this reason, not only does the difference in the flow rates of the sample solution, the washing solvent, and the extraction solvent between the solid phase extraction columns occur, but also the problem arises that the reproducibility of the analysis results is reduced.

  The present invention has been made in view of the situation as described above, and the conditioning operation of the solid phase extraction column can be performed with a small amount of solvent, and the air present around the solid phase adsorbent particles can be obtained. Liquid component analysis method using a solid-phase extraction column that can be replaced with a solvent with high degree of variation, and liquid component analysis assistance that can easily realize such operation An object is to provide an apparatus.

  As a result of intensive studies to solve the above-mentioned problems, the present inventors existed around solid-phase adsorbent particles by performing a conditioning operation by placing the entire solid-phase extraction column in a reduced-pressure environment. The present inventors have found that it is possible to replace air with a solvent quickly and at a high level, thereby completing the present invention.

  The present invention is (1) a liquid component analysis method using a solid phase extraction column having a packed bed filled with a solid phase adsorbent for adsorbing components contained in a liquid sample, At least a conditioning step of supplying a substitution solvent not included to wet the packed bed with the substitution solvent, and placing the entire solid phase extraction column in a reduced pressure environment in the conditioning step, The liquid component analysis method is characterized in that the removal of the air contained in the water is promoted.

  The present invention also includes (2) a packed bed filled with a solid phase adsorbent for adsorbing components contained in the liquid sample, and a liquid for holding the liquid sample above the packed bed. A holding part and an opening for supplying a liquid sample to the liquid holding part are provided, and an outlet for discharging the liquid that has passed through the packed bed is provided below the packed bed. A liquid component analysis auxiliary device used in liquid component analysis using a column, wherein a liquid sample is supplied from the opening to the liquid holding unit inside the liquid component analysis auxiliary device, A working area for collecting necessary liquid flowing out from the outlet, a disposal area for discarding unnecessary liquid flowing out from the outlet, and the solid-phase extraction column standing up Between the work area and the disposal area. And a column mounting table that is movable, and connecting means for connecting an exhaust port for exhausting the air inside the analysis auxiliary device to the outside, on a wall that separates the inside and the outside of the analysis auxiliary device Is a liquid component analysis assisting device.

  According to the present invention, the conditioning operation of the solid-phase extraction column can be performed with a small amount of solvent, and the substitution of the air present around the solid-phase adsorbent particles with the solvent is highly variable with little variation. A liquid component analysis method using a solid phase extraction column, and a liquid component analysis auxiliary device that can easily realize such an operation are provided.

It is a schematic diagram which shows an example of the column used with one embodiment of the analysis method of the liquid component of this invention. It is a schematic diagram which shows an example of the column used by embodiment different from one embodiment of the analysis method of the liquid component of this invention. It is a perspective view showing one embodiment of a liquid component analysis auxiliary device of the present invention.

  Hereinafter, an embodiment of the liquid component analysis method of the present invention will be described. This embodiment is a method for analyzing a liquid component using a column packed with a solid phase adsorbent, and performs a conditioning process in which the column is placed in a reduced pressure environment and the solid phase adsorbent is wetted with a solvent. is there. First, a solid phase extraction column packed with a solid phase adsorbent used in this embodiment will be described.

<Solid phase extraction column>
A solid phase extraction column (hereinafter, also simply referred to as “column”) used in the liquid component analysis method of this embodiment will be described with reference to FIG. FIG. 1 is a schematic diagram showing an example of a column 3 used in the liquid component analysis method of this embodiment. In the column 3, for example, a packed bed 34 filled with a solid-phase adsorbent for adsorbing a sample component from a sample solution is formed inside a hollow body made of a resin such as polyethylene or polypropylene. An opening 31 for supplying a liquid sample and a liquid holding unit 32 for holding the supplied liquid sample are provided above the packed bed 34, and below the packed bed 34, An outlet 36 is provided for allowing the liquid that has passed through the packed bed 34 to flow out.

  The solid phase adsorbent is sandwiched between porous resin sheets 33 and 35 called Fritz and filled in the column 3 to form a packed bed 34. The porous frits 33 and 35 allow liquid to pass through, but do not allow solids to pass through. For this reason, the solid-phase adsorbent particles contained in the packed bed 34 fluctuate in the liquid existing in the liquid holding unit 32 due to the impact when the sample solution is supplied to the column 3 or flow out together with the liquid from the outlet 36. Is prevented. The resin constituting Fritz 33 and 35 is not particularly limited, and examples thereof include polyethylene, polypropylene, polyethylene terephthalate, nylon, and the like. Among these, polyethylene is preferable.

As the solid-phase adsorbent contained in the packed bed 34, an adsorbent usually used in chromatography can be used without particular limitation. Examples of such a solid-phase adsorbent include _C18 silica, silica, porous polymer, carbon and the like. The surface of silica may be modified with an organic compound or the like. Examples of the porous polymer include styrene, divinylbenzene, and methacrylate. These solid-phase adsorbents are packed in the column 3 by a dry method in a powder state, but are filled in the column 3 in a state of being dispersed in a solvent, and then the wet method in which the solvent is removed from the solid-phase adsorbent. The column 3 may be filled with The particle size of the solid phase adsorbent is preferably 10 to 1000 μm, and more preferably 20 to 100 μm.

  The column 3 is stored in a dry state in which the inner packed bed 34 does not contain a solvent. In use, the inner packed bed 34 is wetted with the solvent by passing an appropriate solvent through the column 3.

<Method for analyzing liquid components>
The liquid component analysis method according to this embodiment is performed using the above-described column 3 and has at least a conditioning step of wetting the packed bed 34 with a substitution solvent. After the conditioning step, the target component contained in the liquid sample Or it has the adsorption | suction process which makes a solid-phase adsorbent contained in the packed bed 34 adsorb | suck a non-target component. Hereinafter, each step will be described.

[Conditioning process]
In the conditioning step in the liquid component analysis method of this embodiment, a replacement solvent that does not contain a sample component (hereinafter also referred to as “substitution solvent”) is supplied from the opening 31 of the column 3 to the liquid holding unit 32. This is a step of wetting the packed bed 34 with the replacement solvent by causing the replacement solvent to flow out from the outlet 36 of the column 3.

  This step is a step of wetting the packed bed 34 in a dry state without containing a solvent with a substitution solvent. By passing through this process, the air which exists around the particle | grains of the solid-phase adsorbent contained in the packed bed 34 is substituted by the substitution solvent, and the solid-phase adsorbent is in a state where it can exhibit the adsorption performance.

  The substitution solvent is not particularly limited, and a solvent usually used in chromatography is used. If the solid phase adsorbent contained in the packed bed 34 is for normal phase (the surface of the solid adsorbent particles is hydrophilic), the substitution solvent is, for example, hexane, toluene, chloroform, dichloromethane, benzene, etc. A nonpolar solvent is used. These nonpolar solvents are used alone or in combination of two or more, and are used in a state where polar solvents such as acetone and diethyl ether are mixed as necessary. In addition, if the solid phase adsorbent contained in the packed bed 34 is for reverse phase (the surface of the solid adsorbent particles is hydrophobic), for example, the replacement solvent may be water such as methanol and acetonitrile first. A miscible organic solvent is used, and then water or a buffer (buffer solution), or a mixed solvent in which water or a buffer and one or more organic solvents miscible with the above water are mixed is used.

  By the way, in the conventional liquid component analysis method, as a conditioning process, a process of reducing the pressure on the outlet 36 side of the column 3 and allowing the replacement solvent to pass through the column is generally performed. That is, a pressure difference was provided between the opening 31 of the column 3 and the outlet 36, and the displacement solvent was accelerated by the pressure difference and passed through the packed bed 34 of the column 3. By doing in this way, since a large amount of solvent can be passed through the column 3 in a short time, the time of the conditioning process was shortened.

  However, in such a conventional method, a large amount of substitution solvent is required to replace the air on the surface of the solid phase adsorbent particles contained in the packed bed 34 with the substitution solvent. There was a problem that increased. In addition, since it is not known how much substitution solvent is passed through the column 3 and the air on the surface of the solid-phase adsorbent particles contained in the packed bed 34 is completely substituted with the substitution solvent, the plurality of columns 3 are conditioned. In this case, the degree of substitution by the substitution solvent on the surface of the solid-phase adsorbent particles is different for each column 3, which causes a problem of reducing the reproducibility of the analytical operation. In addition, there is a problem in that a channeling phenomenon due to partial air remaining in the packed bed 34 significantly affects the adsorption capacity of the column 3. Furthermore, when a large amount of solvent is forced to flow inside the column 3 in this way, there is a risk that a large amount of air enters the packed bed 34 as the solvent dies and the column 3 becomes unusable. There is also a problem that the replacement solvent must always be managed so as to remain in the liquid holding part 32 of the column 3.

  In order to solve such problems, the liquid component analysis method of the present embodiment is characterized in that the entire column 3 is placed in a reduced pressure environment in the conditioning step. By having such characteristics, the liquid component analysis method of the present embodiment can solve the problem of conditioning in the conventional liquid component analysis method as described above.

  In the conditioning process, the entire column 3 is placed in a reduced pressure environment as described above, thereby removing air contained in the packed bed 34, that is, air present on the surface of the solid-phase adsorbent particles contained in the packed bed 34. Is promoted. In the conventional method, since the air contained in the packed bed 34 was caused to flow out from the outlet 36 together with the replacement solvent by circulating the replacement solvent, a large amount of solvent was required for the replacement. According to this, the air contained in the packed bed 34 becomes air bubbles by itself when it is placed in a reduced pressure environment and escapes from the packed bed 34. For this reason, the air contained in the packed bed 34 can be removed in a shorter time than the conventional method.

  The conditioning process in the liquid component analysis method of this embodiment will be described more specifically. In the conditioning step in the liquid component analysis method of the present embodiment, a work container with a lid that can be sealed is used. Then, in the state where the column 3 stands up so that the outflow port 36 of the column 3 faces downward inside the working container and there is a space between the working container and the outflow port 36 of the column 3. Standing means for holding is provided.

  The column 3 to be conditioned is held in a standing state on a standing means provided inside the work container, and a substitution solvent is supplied from the opening 31 of the column 3 to the liquid holding unit 32. For example, in the case of a 6 mL tube, for example, the liquid holding unit 32 of the column 3 can normally hold about 5 mL of liquid, so that the substitution solvent is supplied until the liquid holding unit 32 is filled with the substitution solvent.

  The substitution solvent supplied to the liquid holding unit 32 flows out from the outlet 36 of the column 3 due to gravity. However, since the packed bed 34 is filled with the solid-phase adsorbent at a high density and air is present in the packed bed 34, it takes 10 to 60 minutes until all of the substitution solvent supplied to the liquid holding unit 32 flows out. It takes a certain amount of time, and the substitution solvent may not flow out in some cases. Therefore, during the conditioning while reducing the pressure as described below, the replacement solvent is held in the liquid holding unit 32. In addition, it is preferable that a receiving container for receiving the substitution solvent flowing out from the outlet 36 of the column 3 is provided below the column 3.

  As soon as the substitution solvent is supplied to the liquid holding unit 32 of the column 3, the working container is immediately sealed, and the inside of the working container is decompressed. Therefore, the work container is provided with an exhaust port that allows the inside and outside of the work container to communicate with each other and to be connected to an exhaust means (pressure reduction means) provided outside. As the exhaust means provided outside the working container, a known exhaust means can be used without particular limitation, and examples thereof include a water flow pump (aspirator), a vacuum pump and the like.

  The degree of vacuum inside the work container is preferably 600 Torr or more, and more preferably 400 Torr or more. By setting the degree of vacuum inside the working container to 600 Torr or more, the air contained in the packed bed 34 of the column 3 can be efficiently removed. Here, “the degree of decompression inside the working container is 600 Torr or more” means that the pressure inside the working container is 600 Torr or less. In order to prevent the substitution solvent from excessively evaporating or boiling, it is preferable to appropriately adjust the upper limit of the degree of vacuum according to the type of the substitution solvent. For example, a solvent having a low boiling point such as acetone can prevent the displacement solvent from excessively evaporating or boiling by setting the degree of vacuum inside the working container to 180 Torr or less. In order to adjust the degree of decompression inside the work container, it is preferable to provide a pressure regulating valve between the work container and the exhaust means.

  By reducing the pressure inside the working container, the air contained in the packed bed 34 of the column 3 rises as bubbles in the substitution solvent held in the liquid holding unit 32. Further, even under reduced pressure, the solvent flows out from the outlet 36 of the column 3 by gravity, and at this time, the air contained in the packed bed 34 is also removed from the column outlet 36 together with the solvent. The time for maintaining the reduced pressure state varies depending on the amount of the solid-phase adsorbent contained in the packed bed 34, but is preferably about 30 seconds to 5 minutes, more preferably about 30 seconds to 2 minutes, and about 1 minute to 2 minutes. Is most preferred.

  After the elapse of the decompression time, air is introduced into the work container to bring the inside of the work container to normal pressure. Then, the air that has not yet been removed inside the packed bed 34 contracts from the expanded state at the time of decompression as the surrounding environment becomes normal pressure. At that time, the solvent penetrates into the packed bed 34, and the air present around the solid-phase adsorbent particles contained in the packed bed 34 is replaced with the solvent. As a result, although depending on the type of the substitution solvent, the packed layer 34 changes from a white and opaque appearance before the conditioning process to an appearance with increased transparency, and the air contained in the packed layer 34 is removed. I understand that. Thereafter, the degree of air removal inside the packed bed 34 may be confirmed by reducing the pressure again and checking whether or not bubbles are generated from the packed bed 34.

  Thereafter, the substitution solvent remaining in the liquid holding unit 32 of the column 3 is caused to flow out of the outlet 36 of the column 3 by gravity. The conditioning process ends when the replacement solvent remaining in the liquid holding unit 32 finishes flowing out.

[Adsorption process]
The adsorption process is performed using the column 3 that has completed the conditioning process. In the adsorption step in the liquid analysis method of this embodiment, a liquid sample is supplied from the opening 31 of the column 3 to the liquid holding unit 32, and the supplied sample solution is discharged from the outlet 36 of the column 3, thereby In this step, a target component or a non-target component contained in the sample is adsorbed on a solid phase adsorbent contained in the packed bed 34.

  The liquid sample supplied to the liquid holding unit 32 of the column 3 passes through the packed bed 34 and flows out from the outlet 36 of the column 3 by gravity or capillary action. At this time, the target component or non-target component contained in the liquid sample is adsorbed by the solid phase adsorbent contained in the packed bed 34.

  The purpose of using the column 3 in the analysis of liquid is roughly divided into two modes. One is concentration of target components contained in the liquid sample, and the other is removal of non-target components contained in the liquid sample. “Concentration of the target component contained in the liquid sample” means that the target component contained in the liquid sample is adsorbed by the solid phase adsorbent inside the packed bed 34 and then the solvent that causes the target component to flow out is passed through the column 3. It means that the target component contained in the liquid sample is concentrated by removing the target component from the solid phase adsorbent. The “removal of non-target components contained in the liquid sample” means that the non-target components contained in the liquid sample are adsorbed on the solid-phase adsorbent inside the packed bed 34, and the non-target components are discharged from the outlet 36 of the column 3. It is meant to obtain a liquid sample from which water has been removed. In addition, the column 3 is used for the purpose of separating a plurality of components contained in the liquid component by utilizing the difference in the degree of adsorption to the solid phase adsorbent inside the packed bed 34.

  As already described, the liquid sample supplied to the liquid holding part 32 of the column 3 is passed through the packed bed 34 inside the column 3 by gravity or capillary action. At this time, the column 3 is preferably allowed to stand in a standing state. If the column 3 is tilted or vibration is applied to the column 3 while the liquid sample is passed through the packed bed 34, the concentration operation or separation operation may not be performed normally.

  In addition, when the liquid sample supplied to the liquid holding unit 32 of the column 3 flows out from the outlet 36 of the column 3, a thin tube called a disposable liner may be attached to the outlet 36 of the column 3. When a disposable liner is attached to the outlet 36 of the column 3, the flow of the liquid sample from the outlet 36 of the column 3 is accelerated by capillary action.

  When all of the liquid sample supplied to the liquid holding unit 32 of the column 3 flows out from the outlet 36 of the column 3, the adsorption process ends. Thereafter, a liquid sample containing a target component obtained by passing a solvent for causing the target sample to flow out through the packed bed 34 of the column 3, or a liquid from which non-target components have been removed by passing through the packed bed 34 of the column 3. What is necessary is just to perform a required process with respect to a sample suitably and to analyze by a high performance liquid chromatography etc.

According to the liquid component analysis method of the present embodiment, the following effects are exhibited.
In the liquid component analysis method of the present embodiment, the entire column 3 is placed in a reduced pressure environment in the conditioning step of conditioning the column 3. Therefore, the air contained in the packed bed 34 inside the column 3 is quickly removed, and the removal of air from the packed bed 34 is achieved at a higher level than in the conventional method. For this reason, the quantity of the solvent used at a conditioning process can be decreased, and the adsorption | suction performance of the solid-phase adsorbent contained in the packed bed 34 can be exhibited favorably. In addition, since the degree of conditioning of the column 3 can be made uniform, it is possible to reduce variation in analysis results.

  As mentioned above, although one embodiment of the analysis method of the liquid component of the present invention was described, the analysis method of the liquid component of the present invention is not limited to the above embodiment at all, and within the scope of the object of the present invention, Changes can be made as appropriate. For example, in the above embodiment, the syringe type column 3 shown in FIG. 1 is used, but a cartridge type column 3a shown in FIG. 2 may be used. FIG. 2 is a schematic diagram showing an example of a column used in an embodiment different from the embodiment of the liquid component analysis method of the present invention. In this case, a reservoir for supplying a liquid to the liquid holding part 32a of the column 3a is preferably connected to the opening 31a.

<Liquid component analysis aid>
Next, an embodiment of the liquid component analysis auxiliary device of the present invention will be described with reference to FIGS. FIG. 3 is a perspective view showing an embodiment of the liquid component analysis auxiliary device of the present invention.

  The liquid component analysis assisting apparatus 1 of the present embodiment is an apparatus suitably used in the liquid component analysis method of the present invention described above. In the description of the liquid component analysis assisting apparatus 1 of the present embodiment, the description of the portions and effects that overlap with the liquid component analysis method of the present invention described above will be omitted.

  The liquid component analysis auxiliary device 1 of the present embodiment has a container 5 that can be sealed. The inside of the container 5 flows out of the column 3 and a work area 9 for supplying a liquid sample to the liquid holding unit 32 from the opening 31 of the column 3 or collecting a necessary liquid flowing out of the column 3. It is divided into a disposal area 10 for discarding unnecessary liquid. The liquid component analysis performed using the liquid component analysis assisting apparatus 1 of the present embodiment is the same as the column 3 used in one embodiment of the liquid component analysis method of the present invention described above. A column is used.

  The container 5 is provided with a lid 51 at the top. The lid 51 can be opened and closed. For example, the lid 51 is opened when the column 3 is installed on the column installation base 2 or when a liquid is supplied to the column 3. Further, the container 5 is provided with a door 52 in front of the container 5. The door 52 can be opened and closed. For example, the door 52 is opened when the receiving container 7 is installed inside the container 5 or when the column mounting table 2 is moved between the work area 9 and the disposal area 10. The door 52 is preferably made of a transparent material in order to facilitate observation of the inside of the container 5. The container 5 includes a side plate 53, a back plate 54 and a bottom plate 55 in addition to the lid 51 and the door 52.

  The work area 9 is provided with a space for installing a receiving container 7 for receiving the necessary liquid flowing out from the column 3, and the disposal area 10 is used for discarding unnecessary liquid flowing out from the column 3. A waste liquid tank 6 is provided.

  The column mounting table 2 is provided with a plurality of holes for mounting the column 3 in an upright state. Further, the column mounting table 2 is guided by the rail 8 and can move between the work area 9 and the disposal area 10. Therefore, when the liquid is supplied to the liquid holding unit 32 of the column 3 placed on the column setting table 2 or when the necessary liquid flowing out from the outlet 36 of the column 3 is collected, the column setting table 2 is operated. It is possible to move to the area 9 to work, and when discarding the liquid that does not need to flow out of the column 3, the column mounting base 2 is moved to the disposal area 10 and the liquid that has flowed out of the column 3 It can be discarded in the tank 6.

  An exhaust port 4 for connecting an exhaust means 11 for exhausting the air inside the container 5 to the outside is provided on the wall separating the inside and the outside of the container 5. The wall provided with the exhaust port 4 may be any of the lid 51, the door 52, the side plate 53, the back plate 54 and the bottom plate 55. The exhaust port 4 is provided so as to communicate the inside and the outside of the container 5. For this reason, the air inside the container 5 is exhausted to the outside by connecting the exhaust means 11 to the exhaust port 4 with the lid 51 and the door 52 closed and the container 5 sealed, and the inside of the container 5 is decompressed. can do. Under this reduced pressure environment, the column 3 is subjected to the conditioning process described in the embodiment of the liquid component analysis method of the present invention.

  Next, an example of a liquid component analysis method using the liquid component analysis auxiliary device 1 having the above-described configuration will be described. The details of the method for analyzing a liquid component are the same as those of the embodiment of the method for analyzing a liquid component of the present invention described above, so only the outline will be described here.

  First, the lid 51 or the door 52 of the container 5 is opened, and the column 3 is placed on the column installation base 2. The column mounting base 2 is provided with a plurality of holes, but the column 3 may be placed using any of the holes.

  Thereafter, conditioning of column 3 is performed. First, a substitution solvent is injected from the opening 31 provided above the column 3, and the liquid holding part 32 of the column 3 is filled with the substitution solvent. At this time, the column mounting base 2 may be in the work area 9 or the disposal area 10. When the injection of the substitution solvent into the column 3 is completed, the column mounting table 2 is moved to the disposal area 10.

  Next, the lid 51 and the door 52 of the container 5 are closed, and the exhaust means 11 is connected to the exhaust port 4 provided in the container 5 to reduce the pressure inside the container 5. As the inside of the container 5 is depressurized, the air contained in the packed bed 34 inside the column 3 is discharged as bubbles.

  When the air contained in the packed bed 34 inside the column 3 is removed, the exhaust means 11 is removed from the exhaust port 4 to bring the inside of the container 5 to normal pressure. At this time, if the substitution solvent remains in the liquid holding unit 32 of the column 3, the column mounting table 2 may be placed in the disposal area 10 until the substitution solvent in the liquid holding unit 32 becomes empty.

  Next, an adsorption process is performed in which the components in the sample solution are adsorbed by the solid phase adsorbent contained in the packed bed 34. The door 52 of the container 5 is opened, and a receiving container 7 for collecting a liquid sample is disposed at the bottom of the work area 9. The receiving container 7 is disposed so as to be positioned below the column 3 when the column mounting base 2 moves to the work area 9. Then, the sample solution is supplied to the liquid holding unit 32 of the column 3.

  As already described, the component adsorbed by the solid-phase adsorbent inside the packed bed 34 may be a component necessary for the subsequent analysis operation or an unnecessary component. If the component adsorbed on the solid-phase adsorbent is a necessary component, the component flowing out from the outlet 36 of the column 3 is unnecessary, so the adsorption step is performed in a state where the column installation base 2 exists in the disposal area 10. Just do it. Thereby, an unnecessary outflow component flows out into the waste liquid tank 6. On the other hand, when the component adsorbed on the solid-phase adsorbent is an unnecessary component, the component that flows out from the outlet 36 of the column 3 is necessary, so that the adsorption process is performed in a state where the column installation base 2 exists in the work area 9. Can be done. Thereby, the necessary outflow component flows out into the receiving container 5. In any case, when the sample supplied to the liquid holding unit 32 of the column 3 is emptied, a solvent is appropriately added to sufficiently flow out unnecessary components or necessary components.

  When the component adsorbed on the solid phase adsorbent in the packed bed 34 in the adsorption step is a necessary component, the component adsorbed on the solid phase adsorbent needs to be discharged to the container 5. For this purpose, the elution solvent is supplied to the column 3 in a state where the column mounting base 2 exists in the work area 9. The elution solvent is a solvent for eluting the target component adsorbed on the solid phase adsorbent, and is appropriately selected according to the type of the solid phase adsorbent and the target component adsorbed on the solid phase adsorbent. Is.

  As a result of the above operations, the solution containing the target component is stored in the receiving container 5, and thereafter, concentration may be appropriately performed or analysis may be performed by high performance liquid chromatography or the like.

As mentioned above, although one embodiment of the liquid component analysis auxiliary device 1 of the present invention has been described, the liquid component analysis auxiliary device 1 of the present invention is not limited to the above embodiment, and the object of the present invention. Within the range, it can implement by changing suitably.
For example, in the above embodiment, the lid 5 and the door 52 are provided on the container 5, but only one of these may be provided, and even more doors and lids may be provided. Good. Moreover, in the said embodiment, although the rail 8 was provided as a guide for moving the column installation base 2 manually, another guide means may be used and the movement means may not be manual. .

  EXAMPLES Hereinafter, although the analysis method of the liquid component of this invention is demonstrated in detail by an Example, the following Examples illustrate this invention and this invention is not limited to the following Examples at all.

[Example 1]
A column mounting table was provided inside the sealable container, and a column for solid phase extraction (ENVI-Carb / LC-NH ₂ = 500 mg / 500 mg / 6 mL, manufactured by Sigma-Aldrich) was mounted on the column mounting table. Thereafter, as a conditioning step, 5 mL of a substitution solvent consisting of acetonitrile: toluene = 3: 1 was supplied to the liquid holding unit at the top of the column, and the inside of the sealed container was depressurized to place the entire column in a reduced pressure environment. The degree of decompression inside the sealed container at this time was 250 Torr, and the decompressed state was maintained for 1 minute. Thereafter, the reduced pressure inside the sealed container was released to normal pressure and left for 30 seconds. In order to confirm that the air of the solid-phase adsorbent in the column has been released, the inside of the sealed container is again depressurized (250 Torr), and after confirming that no bubbles are generated, the inside of the sealed container is again kept inside. Returned to pressure. During the conditioning process, the displacement solvent supplied to the liquid holding part of the column continues to flow out from the outlet provided in the lower part of the column due to gravity and capillary action, and the above-described decompression-normal pressure operation is completed. The conditioning process was terminated when 5 mL of the replacement solvent supplied to the liquid holding part of the column became empty. Therefore, the displacement solvent used in the conditioning process in Example 1 is 5 mL.

  A 25 mL reservoir for supplying liquid to the liquid holding part of the column was attached to the upper part of the column that was conditioned by the above process, and a disposable liner was attached to the outlet of the column. Thereafter, 10 mL of a solvent consisting of acetonitrile: toluene = 3: 1 was added to the reservoir, and the time required for the solvent to pass through the inside of the column due to natural fall was measured. This measurement was repeated 10 times, and an average value, a standard deviation, and a coefficient of variation were calculated from the experimental results for 8 times excluding the largest value and the smallest value. The results are shown in Table 1.

[Example 2]
The test was performed under the same conditions as in Example 1 except that the amount of the displacement solvent used in the conditioning process was changed to 3 mL. The results are shown in Table 2.

[Comparative Example 1]
A solid-phase extraction column is attached to a vacuum manifold (an apparatus that allows liquid to pass through the column at a high speed by reducing the pressure at the outlet of the solid-phase extraction column). A 25 mL reservoir to supply the liquid was attached. By supplying 10 mL of a displacement solvent consisting of acetonitrile: toluene = 3: 1 to the liquid holding part of the column via the reservoir, operating the vacuum manifold and reducing the outlet of the column to a reduced pressure (630 Torr), The conditioning process was performed by flowing out 10 mL of the supplied substitution solvent. After the displacement solvent level was close to the frits at the top of the column, the vacuum manifold valve was closed to stop the displacement solvent outflow and the conditioning process was completed. Therefore, the displacement solvent used for the conditioning process in Comparative Example 1 is 10 mL.

  A 25 mL reservoir for supplying a liquid to the liquid holding part of the column was attached to the upper part of the column that was conditioned by the above process, and a disposable liner was attached to the outlet of the column. Thereafter, 10 mL of a solvent consisting of acetonitrile: toluene = 3: 1 was added to the reservoir, and the time required for the solvent to pass through the inside of the column due to natural fall was measured. This measurement was repeated 10 times, and an average value, a standard deviation, and a coefficient of variation were calculated from the experimental results for 8 times excluding the largest value and the smallest value. The results are shown in Table 3.

The liquid outflow time (flow rate) of Comparative Example 1 (Table 3) was 18 minutes 23 seconds (0.6 mL / min), whereas in Example 1 (Table 1) and Example 2 (Table 2). The liquid outflow time (flow rate) was 7 minutes 42 seconds (1.4 mL / min) and 9 minutes 16 seconds (1.1 mL / min), respectively. Therefore, the column subjected to the conditioning process in the liquid component analysis method of the present invention has a liquid flow rate twice or more faster than the column not subjected to the conditioning process in the liquid component analysis method of the present invention. It is understood that the air contained in the packed bed of the column has been removed at a higher level.
In addition, the variation coefficients of Example 1 (Table 1) and Example 2 (Table 2) are both smaller than those of Comparative Example 1 (Table 3), and the conditioning process in the liquid component analysis method of the present invention is employed. By doing so, it can be seen that the variation in the liquid outflow time of the column after conditioning becomes small. In other words, it can be seen that by adopting the conditioning process according to the present invention, the degree of variation in the columns after conditioning can be reduced. Since the liquid outflow time of the column is a factor that greatly affects the adsorption of the sample components to the solid phase adsorbent of the column, by adopting the liquid component analysis method of the present invention, it is Analysis can be performed. Further, in Example 2 (Table 2), the amount of the substitution solvent used for conditioning was 3 mL, and although the amount of substitution solvent used was 30% compared to Comparative Example 1 (Table 3), the comparison was made. The degree of variation in the column after conditioning is smaller than in Example 1. From this, it was shown that the amount of the substitution solvent used in the conditioning process can be reduced by employing the liquid component analysis method of the present invention.

DESCRIPTION OF SYMBOLS 1 Liquid component analysis assistance apparatus 2 Column installation stand 3 Column for solid phase extraction 31 Opening part 32 Liquid holding part 34 Packing layer 36 Outlet 4 Exhaust port 5 Container 6 Waste liquid tank 7 Receiving container 8 Rail 9 Work area 10 Waste area 11 Exhaust means

Claims (2)

A liquid component analysis method using a solid phase extraction column having a packed bed filled with a solid phase adsorbent for adsorbing components contained in a liquid sample,
At least a conditioning step of supplying a substitution solvent not containing a sample component to wet the packed bed with the substitution solvent;
In the conditioning step, the entire solid phase extraction column is placed in a reduced pressure environment to promote the removal of air contained in the packed bed, and the liquid component analysis method is characterized in that: Having a packed bed filled with a solid phase adsorbent for adsorbing components contained in a liquid sample;
Above the packed bed, a liquid holding unit for holding the liquid sample and an opening for supplying the liquid sample to the liquid holding unit are provided,
Below the packed bed is a liquid component analysis auxiliary device used in the analysis of the liquid component using a solid phase extraction column provided with an outlet for flowing out the liquid that has passed through the packed bed,
Inside the liquid component analysis aid device,
A working area for supplying a liquid sample from the opening to the liquid holding unit or collecting a necessary liquid flowing out from the outlet,
A disposal area for discarding unnecessary liquid that has flowed out of the outlet;
The solid phase extraction column is installed upright, and a column installation table that is movable between the work area and the disposal area is provided,
A liquid component analysis auxiliary device, wherein an exhaust port for connecting an exhaust means for exhausting the air inside the analysis auxiliary device to the outside is provided on a wall separating the inside and the outside of the analysis auxiliary device.

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