JP6143528B2 - Mercury analysis pretreatment method, mercury analysis pretreatment device, mercury analysis method, mercury analysis device, and mercury desorption solution - Google Patents

Description

  The present invention relates to a mercury analysis pretreatment method, a mercury analysis pretreatment apparatus, a mercury analysis method, a mercury analysis apparatus, and a mercury desorption solution used for these.

  Sources of mercury released into the environment are broadly classified into two categories: natural activities such as volcanic activity and rock weathering, and human activities. There are many sources of mercury derived from human activities, such as mining and burning fossil fuels such as coal, metal production including gold and iron, cement production, and waste combustion.

  In recent years, due to global pollution concerns, the United Nations Environment Program (UNEP) has been debating international efforts related to mercury consumption, disposal, and movement between countries. From 2010, an intergovernmental bargaining conference was started to prepare a global legal framework for mercury, and the work results are scheduled to be reported to the 27th UNEP Executive Board meeting in 2013. The Diplomatic Conference will adopt a treaty that sets certain restrictions on the export, import, use and disposal of mercury.

  Against this background, it is expected that there will be a need for mercury analysis that promptly determines whether standards have been exceeded at various stages from the export, import, production, and disposal of materials. Therefore, in addition to the conventional instrumental analysis and the existing simple measurement, there is a demand for a measurement method that enables mercury measurement more easily, quickly, and inexpensively.

  Until now, instrumental analysis methods such as atomic absorption spectrometry have been used as mercury measurement methods. These methods require expensive equipment, require a certain amount of time for analysis, and in principle, analysis must be performed in a laboratory. In recent years, as a simple analysis method for mercury, a method based on an electrochemical measurement method typified by a stripping method (for example, see Non-Patent Document 1) or a fluorescent X-ray analysis (for example, see Non-Patent Document 2) has been used. Has been developed. However, there is a problem that both require expensive introduction costs. In addition, a simple measurement using a DNA aptamer (for example, see Non-Patent Document 3) has been proposed as a new technique, but this is not a technique until practical use.

  On the other hand, for the purpose of application to environmental measurement, the present applicant has developed an antibody that specifically binds heavy metal and a highly sensitive flow immunoassay using the antibody (for example, non-patent literature). 4 and 5). Therefore, it is conceivable to apply these techniques to mercury measurement methods. However, in the sample where mercury coexists with various metals, the problem is that the quantitativeness is hindered by the influence of other metals and the measurement accuracy is lowered. There is.

  Therefore, in order to increase the measurement accuracy of mercury determination in a measurement method such as an immunoassay, a pretreatment for removing other metals is necessary. Further, it is desired that the pretreatment can be performed simply, quickly, and inexpensively.

  Therefore, for example, an adsorption step in which a sample containing a heavy metal to be extracted is adsorbed to the cation exchange resin using a cation exchange resin, and a removal step in which heavy metals not to be extracted are removed from the cation exchange resin by washing. And a heavy metal extraction method that includes an elution step of eluting a heavy metal to be extracted from the cation exchange resin (see, for example, Patent Document 1). However, the proposed technique requires elution using a relatively high concentration of acid, and is problematic in that it is difficult to use a simple method.

  Therefore, a mercury analysis pretreatment method, a mercury analysis pretreatment device, a mercury analysis method, a mercury analysis device, and a mercury used in these, which can be used for a quantitative measurement method of mercury and can be performed simply, quickly and inexpensively. At present, it is required to provide a desorption solution.

JP 2009-294024 A

C. Chen, J.A. Zhang, Y. et al. Du, X. Yang, E .; Wang, Analyst, 135 (5), 1010-1014 (2010) D. L. Anderson, J AOAC Int. Mar-Apr 93 (2), 683-93 (2010) Z. Jiang, Y.J. Fan, M.M. Chen, A.M. Liang, X.M. Liao, G.G. Wen, X. Shen, X. He, H. Pan, and H.C. Jiang, Anal. Chem. 81, 5439-5445 (2009) N. Ohmura, S .; J. et al. Lackie, H.M. Saiki: Anal. Chem. , 73, 3392-3399 (2001) T. T. R. Glass, H.M. Saiki, T .; Joh, Y. Taemi, S.M. J. et al. Lackie, N.M. Ohmura: Biosensors and Bioelectronics, 20, 397-403 (2004)

  An object of the present invention is to solve the above-described problems and achieve the following objects. That is, the present invention is a mercury analysis pretreatment method, a mercury analysis pretreatment device, a mercury analysis method, a mercury analysis device, and a mercury analysis pretreatment method that can be used in a quantitative measurement method for mercury and can be performed simply, quickly, and inexpensively. It aims at providing the mercury desorption solution used for these.

Means for solving the problems are as follows. That is,
The mercury analysis pretreatment method of the present invention is a mercury analysis pretreatment method for separating the mercury and the metal other than mercury from a sample solution containing mercury and a metal other than mercury, wherein the sample solution is treated with an anion. An immobilization step of passing through the exchange resin and immobilizing at least the mercury on the anion exchange resin, and subsequently washing the anion exchange resin with washing water, and removing the metal other than mercury from the anion exchange A washing step for removing from the resin, and subsequently passing a mercury desorption solution for desorbing the mercury from the anion exchange resin through the anion exchange resin to desorb the mercury from the anion exchange resin. And a desorption step.
The mercury analysis pretreatment apparatus of the present invention is a mercury analysis pretreatment apparatus for separating mercury and a metal other than mercury from a sample solution containing mercury and a metal other than mercury, wherein the sample solution is treated with an anion. Immobilization means for passing through an exchange resin and fixing at least the mercury to the anion exchange resin, and washing the anion exchange resin with washing water, and removing metals other than the mercury from the anion exchange resin And a desorption means for passing a mercury desorption solution for desorbing the mercury from the anion exchange resin through the anion exchange resin and desorbing the mercury from the anion exchange resin. It is characterized by that.
The mercury desorption solution of the present invention is characterized by being used for desorbing mercury from an anion exchange resin containing a buffer and immobilizing mercury.
The mercury analysis method of the present invention includes the mercury analysis pretreatment method of the present invention and a quantitative process for quantifying mercury separated from a metal other than mercury by the mercury analysis pretreatment method.
The mercury analyzer of the present invention comprises the mercury analysis pretreatment apparatus of the present invention and a quantitative means for quantifying mercury separated from a metal other than mercury by the mercury analysis pretreatment apparatus.

  According to the present invention, it is possible to solve the above-mentioned problems in the prior art, achieve the above-mentioned object, and can be used for a mercury quantitative measurement method, which can be performed simply, quickly and inexpensively. It is possible to provide a method, a mercury analysis pretreatment apparatus, a mercury analysis method, a mercury analysis apparatus, and a mercury desorption solution used for these.

FIG. 1A is a schematic diagram illustrating an example of how the anion exchange resin is used. FIG. 1B is a schematic diagram for explaining an example of the liquid passing process. FIG. 1C is a schematic diagram for explaining an example of the immobilization process. FIG. 1D is a schematic diagram for explaining an example of the immobilization process. FIG. 1E is a schematic diagram for explaining an example of a cleaning process. FIG. 1F is a schematic diagram for explaining an example of a desorption process. FIG. 2 is a schematic diagram for explaining an example of the immobilization process of the present invention. FIG. 3A is a graph showing the immobilization ratio of mercury to an anion exchange resin when 500 μL of an anion exchange resin having a large particle size is used. FIG. 3B is a graph showing the immobilization ratio of mercury to an anion exchange resin when 100 μL of an anion exchange resin having a large particle size is used. FIG. 3C is a graph showing the immobilization ratio of mercury on an anion exchange resin when 500 μL of an anion exchange resin having a small particle diameter is used. FIG. 3D is a graph showing the immobilization ratio of mercury to an anion exchange resin when 100 μL of an anion exchange resin having a small particle size is used. FIG. 4 is a graph showing the immobilization rate of each metal ion at each hydrochloric acid concentration. FIG. 5A is a graph showing the cleaning rate of iron ions and cadmium ions in the cleaning step when a sample solution having a hydrochloric acid concentration of 0.1 mM is used. FIG. 5B is a graph showing the cleaning rate of iron ions and cadmium ions in the cleaning step when a sample solution having a hydrochloric acid concentration of 0.1 M is used. FIG. 5C is a graph showing the cleaning rate of zinc ions in the cleaning step when a sample solution having a hydrochloric acid concentration of 1M is used. FIG. 6A is a graph showing the mercury desorption rate in the desorption step when a 50 mM Tris buffer is used as the mercury desorption solution. FIG. 6B is a graph showing the mercury desorption rate in the desorption step when 40 mM nitric acid is used as the mercury desorption solution. FIG. 6C is a graph showing the mercury desorption rate in the desorption step when 40 mM sulfuric acid is used as the mercury desorption solution. FIG. 6D is a graph showing the mercury desorption rate in the desorption step when EDTA-4Na 50 mM Tris buffer (pH adjustment with HCl) is used as the mercury desorption solution. FIG. 6E is a graph showing the mercury desorption rate in the desorption step when EDTA-4NH ₄ 50 mM Tris buffer (pH adjustment with HCl) is used as the mercury desorption solution. FIG. 6F is a graph showing the mercury desorption rate in the desorption step when EDTA-4Na 50 mM Tris buffer (pH adjustment with HNO ₃ ) is used as the mercury desorption solution. FIG. 6G is a graph showing the mercury desorption rate in the desorption step when EDTA-4NH ₄ 50 mM Tris buffer (pH adjustment with HNO ₃ ) is used as the mercury desorption solution. FIG. 7A is a graph showing the desorption rate of metal ions to EDTA-4Na 50 mM Tris buffer (pH adjustment with HNO ₃ ) when a sample solution having a hydrochloric acid concentration of 0.1 mM is used. FIG. 7B is a graph showing the desorption rate of metal ions into EDTA-4Na 50 mM Tris buffer (pH adjustment with HNO ₃ ) when a sample solution having a hydrochloric acid concentration of 0.1 M is used. FIG. 8A is a schematic diagram of a microchannel used in the determination step. FIG. 8B is an enlarged view of the measurement site in FIG. 8A. FIG. 8C is a schematic diagram for explaining the principle of microimmunoassay. FIG. 9 is a graph showing the relationship between the mercury concentration and the relative signal value obtained by the quantitative process 1 and the quantitative process 2. FIG. 10A is a graph showing the mercury desorption rate in the desorption step when EDTA-4Na 50 mM Tris buffer (pH adjustment with H ₂ SO ₄ ) is used as the mercury desorption solution. FIG. 10B is a graph showing the mercury desorption rate in the desorption step when EDTA-4Na 50 mM Tris buffer (pH adjustment with H ₃ PO ₄ ) is used as the mercury desorption solution. FIG. 10C is a graph showing the mercury desorption rate in the desorption step when EDTA-4Na 100 mM Tris buffer (pH adjustment with HNO ₃ ) is used as the mercury desorption solution. FIG. 10D is a graph showing the mercury desorption rate in the desorption step when EDTA-4Na 500 mM Tris buffer (pH adjustment with HNO ₃ ) is used as the mercury desorption solution. FIG. 10E is a graph showing the mercury desorption rate in the desorption step when using phosphate buffered saline (PBS) -EDTA-4Na as the mercury desorption solution. FIG. 11 is a schematic view showing a container containing an anion exchange resin and a suction member. FIG. 12 is a graph showing the relationship between pressure and liquid flow rate when a suction member is used. FIG. 13 is a rough diagram showing the relationship between the liquid flow rate and the mercury immobilization rate when a suction member is used. FIG. 14 is a graph showing the relationship between the liquid flow rate and the mercury desorption rate when a suction member is used. FIG. 15 is a schematic view showing a container containing an anion exchange resin, a suction member, and a syringe. FIG. 16 is a graph showing the relationship between the pressure and the flow rate when a suction member is used. FIG. 17 is a graph showing the relationship between the liquid flow rate and the mercury immobilization rate when a suction member is used. FIG. 18 is a graph showing the relationship between the liquid flow rate and the mercury desorption rate when a suction member is used. FIG. 19 is a graph showing the relationship between the amount of the sample solution, the mercury concentration in the sample solution, and the mercury desorption rate. FIG. 20 is a graph showing the relationship between the mercury concentration of the sample solution and the relative signal value. FIG. 21 is a graph showing the relationship between the mercury concentration in the sample solution and the relative signal value in the analysis of the river water sample. FIG. 22 is a graph showing the relationship between the mercury concentration in the sample solution and the relative signal value in the analysis of the soil sample. FIG. 23 is a graph showing the relationship between the mercury concentration in the sample solution and the relative signal value in the analysis of the coal ash sample.

(Mercury analysis pretreatment method, mercury analysis pretreatment device, and mercury desorption solution)
The mercury analysis pretreatment method of the present invention includes at least an immobilization step, a washing step, and a desorption step, and further includes other steps as necessary.
The mercury analysis pretreatment method is a pretreatment method for separating mercury and a metal other than mercury from a sample solution containing mercury and a metal other than mercury.
The mercury analysis pretreatment apparatus of the present invention has at least an immobilization unit, a cleaning unit, and a desorption unit, and further includes other units as necessary.
The mercury analysis pretreatment apparatus is a pretreatment apparatus that separates the mercury and the metal other than mercury from a sample solution containing mercury and a metal other than mercury.
The mercury desorption solution of the present invention contains at least a buffer, and further contains other components as necessary.
The mercury desorption solution is used to desorb mercury from an anion exchange resin on which mercury is immobilized.
The mercury analysis pretreatment method can be preferably carried out by the mercury analysis pretreatment apparatus, the immobilization step can be suitably carried out by the immobilization means, and the washing step can be suitably carried out by the washing means, The desorption step can be preferably performed by the desorption means.

<Immobilization process and immobilization means>
The immobilization step is not particularly limited as long as it is a step of passing a sample solution through an anion exchange resin and immobilizing at least the mercury on the anion exchange resin, and is appropriately selected depending on the purpose. be able to.
The immobilization means is not particularly limited as long as it is a means for passing the sample solution through the anion exchange resin and immobilizing at least the mercury on the anion exchange resin, and is appropriately selected according to the purpose. be able to.

-Sample solution-
The sample solution is not particularly limited as long as it is a solution containing mercury and a metal other than mercury, and can be appropriately selected according to the purpose.
There is no restriction | limiting in particular as metals other than said mercury, According to the objective, it can select suitably, For example, iron, cadmium, etc. are mentioned.
In the present specification, the mercury and the metal may be mercury and a metal that are not in an ionic state, or may be a metal in an ionic state, that is, a mercury ion and a metal ion.

The sample solution preferably contains hydrochloric acid. There is no restriction | limiting in particular as the density | concentration of the said hydrochloric acid in the said sample solution, Although it can select suitably according to the objective, 0.1 mM (mmol / L) -1M (mol / L) is preferable and 1 mM-1M is preferable. More preferably, 5 mM to 0.5 M is still more preferable, and 10 mM to 0.1 M is particularly preferable. Within the preferred range, the immobilization rate of metals other than mercury and cadmium becomes low, and when the concentration is within the further preferred range and the particularly preferred range, the effect becomes remarkable. Cadmium can be easily removed by a washing process, so even if the immobilization rate is high, there is almost no influence on the accuracy of mercury analysis.
If the hydrochloric acid concentration is less than 0.1 mM, iron may not be completely removed in the washing process, and iron may be desorbed together with mercury in the desorption process. Therefore, when the hydrochloric acid concentration is less than 0.1 mM, the combination with other pretreatment capable of removing iron and the selection of an analysis method that does not affect the presence of iron in the mercury analysis after this pretreatment are preferable. .

  The sample solution may be collected from any place, for example, from factory wastewater, waste after incineration, river water, soil, waste containing coal ash, food, organisms, biological samples, etc. Examples include collected sample solutions.

-Anion exchange resin-
The material, shape, size, and structure of the anion exchange resin are not particularly limited and may be appropriately selected depending on the purpose.
Examples of the shape of the anion exchange resin include beads, fibers, and membranes. Among these, a bead shape is preferable in terms of excellent filling properties.
When the anion exchange resin is in the form of beads, the size is not particularly limited and can be appropriately selected according to the purpose. However, the particle size distribution is 10 μm to 200 μm in terms of excellent filling properties. It is preferable.
The anion exchange resin preferably has a quaternary ammonium base.
The anion exchange resin may be a commercially available product. The commercially available product is not particularly limited and may be appropriately selected depending on the purpose, for ^{example, Dowex TM 1x2 100-200, Dowex TM} 1x2 200-400 ( Wako Pure Chemical in both the production of The Dow Chemical Company Can be purchased from Kogyo Co., Ltd.).

  The anion exchange resin is preferably contained in a container.

  Hereinafter, various preferable numerical ranges described in the present specification are particularly preferable ranges when the volume occupied by the anion exchange resin in the container is 100 μL when the anion exchange resin is packed in the container. .

--Container--
There is no restriction | limiting in particular as said container, According to the objective, it can select suitably, For example, a syringe etc. are mentioned. There is no restriction | limiting in particular as a magnitude | size of the said syringe, According to the objective, it can select suitably.

  In the immobilization step and the immobilization unit, the anion exchange resin is, for example, put the anion exchange resin in the container, and the anion exchange resin from both ends of a syringe space containing the anion exchange resin. Is sandwiched between liquid permeable materials such as filter paper.

  Further, in the immobilization step and the immobilization means, the anion exchange resin is preferably filled in a container 1 as shown in FIG. That is, the anion exchange resin is preferably filled in a container so that the anion exchange resin does not move when filled. By doing so, the layer thickness of the anion exchange resin can be made uniform.

-Liquid flow-
The liquid passing method is not particularly limited and may be appropriately selected depending on the purpose. For example, a predetermined amount of the sample solution is passed through a container filled with a predetermined amount of the anion exchange resin. The method etc. are mentioned.
The amount of the sample solution to be passed is not particularly limited and may be appropriately selected depending on the purpose, but is preferably 0.5 mL to 100 mL, and more preferably 2 mL to 100 mL. When the amount of the sample solution to be passed is within the more preferable range, it is advantageous in that a highly accurate mercury analysis measurement can be performed with a small amount of sample.
The liquid passing may be performed once or may be performed in a plurality of times. For example, a total of 50 mL may be performed by passing 5 mL through 10 times.
When the amount of the sample solution is large, for example, in the case of 30 to 100 mL, the amount of the mercury desorption solution in the desorption step is preferably 1/2 to 1 with respect to the amount (mL) of the sample solution. By setting the ratio to / 20, more preferably 1/7 to 1/15, the mercury in the sample solution can be concentrated and used for the determination step of mercury analysis, and the sensitivity of mercury analysis can be improved.
There is no restriction | limiting in particular as a speed | rate of liquid flow, Although it can select suitably according to the objective, 0.1 mL / min-50 mL / min are preferable, 0.2 mL / min-20 mL / min are more preferable, 0 3 mL / min to 10 mL / min is more preferable, and 2.0 mL / min to 6.0 mL / min is particularly preferable.
Yes.
The liquid flow can be performed using a sample solution supply member. Examples of the sample solution supply member include a syringe.

The passing of the sample solution is preferably performed using a suction member. By using the suction member, the liquid passing speed can be increased as compared with the case where liquid is passed without using the suction member (when the liquid is passed naturally). Further, by using the suction member, the liquid passing speed can be controlled with high accuracy.
There is no restriction | limiting in particular as said suction member, According to the objective, it can select suitably, For example, a suction pump etc. are mentioned. Examples of the suction pump include a diaphragm pump and a plunger pump.

Through the immobilization step, the mercury in the sample solution is immobilized on the anion exchange resin.
The mercury in the sample solution is immobilized on the anion exchange resin by the immobilization means.
In the immobilization step, a metal other than the mercury may be immobilized on the anion exchange resin. The immobilized metal other than mercury is removed from the anion exchange resin by a cleaning process described later.

<Washing process and cleaning means>
The washing step is not particularly limited as long as it is a step of washing the anion exchange resin with washing water following the immobilization step and removing metals other than mercury from the anion exchange resin, It can select suitably according to the objective, For example, it can carry out using a pure water.
The washing means is not particularly limited as long as it is a means for washing the anion exchange resin with washing water and removing metals other than mercury from the anion exchange resin, and it is appropriately selected according to the purpose. Can do.

-Washing water-
There is no restriction | limiting in particular as said washing water, Although it can select suitably according to the objective, Pure water is preferable. By using pure water as the washing water, metals other than mercury can be removed without almost desorbing the mercury from the anion exchange resin.

The cleaning method is not particularly limited and can be appropriately selected depending on the purpose. For example, a method of passing a predetermined amount of the cleaning water through a container filled with a predetermined amount of the anion exchange resin, etc. Is mentioned.
There is no restriction | limiting in particular as the quantity of the said washing water in the case of washing | cleaning, Although it can select suitably according to the objective, 1 mL-10 mL are preferable in 1 washing | cleaning, and 3 mL-7 mL are more preferable.
There is no restriction | limiting in particular as the frequency | count of washing | cleaning in the case of washing | cleaning, Although it can select suitably according to the objective, 1 to 3 times is preferable.
There is no restriction | limiting in particular as the speed | rate of liquid flow in the case of washing | cleaning, Although it can select suitably according to the objective, 0.1 mL / min-50 mL / min are preferable, 0.2 mL / min-20 mL / min are preferable. More preferably, 0.3 mL / min to 10 mL / min is more preferable, and 2.0 mL / min to 8.0 mL / min is particularly preferable.
Cleaning can be performed using a cleaning liquid supply member. Examples of the cleaning liquid supply member include a syringe.

The washing water is preferably passed using a suction member. By using the suction member, the liquid passing speed can be increased as compared with the case where liquid is passed without using the suction member (when the liquid is passed naturally). Further, by using the suction member, the liquid passing speed can be controlled with high accuracy.
There is no restriction | limiting in particular as said suction member, According to the objective, it can select suitably, For example, a suction pump etc. are mentioned. Examples of the suction pump include a diaphragm pump and a plunger pump.

  Through the washing step, metals other than mercury can be removed from the anion exchange resin.

<Desorption step and desorption means>
The desorption step is not particularly limited as long as it is a step of passing a mercury desorption solution through the anion exchange resin and desorbing the mercury from the anion exchange resin following the washing step. Can be appropriately selected according to the purpose.
The desorption means is not particularly limited as long as it is a means for passing a mercury desorption solution through the anion exchange resin and desorbing the mercury from the anion exchange resin, and is appropriately selected according to the purpose. can do.

-Mercury desorption solution-
The mercury desorption solution is not particularly limited as long as it is a solution that desorbs the mercury from the anion exchange resin, and can be appropriately selected according to the purpose, but preferably contains a buffer. It is more preferable to contain a chelating agent that chelates the mercury to form a mercury complex and the buffer. When the mercury desorption solution contains the buffer, when an antibody is used for mercury analysis, mercury analysis can be performed without adjusting pH. When the mercury desorption solution contains the chelating agent and the buffering agent, that is, is a chelating agent-containing buffer solution, when an antibody is used for mercury analysis, pH adjustment or a new chelating agent is added. In addition, mercury analysis becomes possible by mixing with antibody.
The mercury desorption solution preferably contains at least one of nitric acid, sulfuric acid and phosphoric acid, and more preferably contains at least one of nitric acid and sulfuric acid. The concentration of the nitric acid, the sulfuric acid and the phosphoric acid in the mercury desorption solution is not particularly limited and can be appropriately selected according to the purpose. However, in terms of desorption ability, 0.1 mM to 1, 000 mM is preferable, 1 mM to 500 mM is more preferable, and 10 mM to 100 mM is particularly preferable.

--Buffer agent--
The buffer is not particularly limited and may be appropriately selected depending on the intended purpose. For example, Good's buffer, phosphate buffer, Tris buffer (trishydroxymethylaminomethane), phosphate buffered physiological saline Water etc. are mentioned.

  The concentration of the buffer in the mercury desorption solution is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 50 mM or more and 50 mM to 1, from the viewpoint of excellent mercury desorption ability. 000 mM is preferable, and 50 mM to 500 mM is more preferable.

  There is no restriction | limiting in particular as a density | concentration of the said buffering agent in the said mercury desorption solution, According to the objective, it can select suitably.

--Chelating agent--
The chelating agent is not particularly limited as long as it chelates the mercury to form a mercury complex, and can be appropriately selected according to the purpose. However, diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid ( EDTA), tetraethylenetriamine (TET), ethylenediamine (EDA), diethylenetriamine (DETA), citric acid, oxalic acid, crown ether, nitrilotetraacetic acid, disodium edetate, sodium edetate, trisodium edetate, penicillamine, pente Tate calcium trisodium, pentetate, succimer, and trientine edetate are preferred, and ethylenediaminetetraacetic acid (EDTA) is more preferred.

  There is no restriction | limiting in particular as a density | concentration of the said chelating agent in the said mercury elimination | desorption solution, Although it can select suitably according to the objective, 0.1 micromol-10 mM are preferable, 0.5 micromol-500 micromol are more preferable, 1 micromol- 100 μM is particularly preferred. When the concentration is within the particularly preferred range, it is advantageous in that mercury after desorption can be stably present in the solution and immunoassay can be performed with high sensitivity.

The pH of the mercury desorption solution when the mercury desorption solution contains the buffer is not particularly limited and may be appropriately selected depending on the intended purpose, but is preferably 1.0 to 8.5. 5.0 to 8.0 is more preferable, and 6.5 to 7.5 is particularly preferable. When the pH is within the preferred range, it is advantageous in that the dissolved mercury (Hg ²⁺ ) is stable, and when the pH is within the particularly preferred range, the pH is not further adjusted. This is advantageous in that it can be used for immunoassays using antibodies.
The method for adjusting the pH is not particularly limited and may be appropriately selected depending on the intended purpose. However, it is adjusted with at least one of nitric acid, sulfuric acid, phosphoric acid and hydrochloric acid in terms of excellent mercury desorption ability. It is preferable to adjust with at least one of nitric acid, sulfuric acid and phosphoric acid.

The amount of the mercury desorbing solution is not particularly limited and may be appropriately selected depending on the intended purpose. However, 0.5 mL to 15 mL is preferable and 2.0 mL to 7.5 mL is preferable in one flow. More preferred.
There is no restriction | limiting in particular as the frequency | count of liquid passage, Although it can select suitably according to the objective, 1 time or more are preferable, 2-5 times are more preferable, and 2 times are especially preferable.
There is no restriction | limiting in particular as a speed | rate of liquid flow, Although it can select suitably according to the objective, 0.1 mL / min-50 mL / min are preferable, 0.2 mL / min-20 mL / min are more preferable, 0 3 mL / min to 10 mL / min is more preferable, and 2.0 mL / min to 8.0 mL / min is particularly preferable.

The passing of the mercury desorbing solution is preferably performed using a suction member. By using the suction member, the liquid passing speed can be increased as compared with the case where liquid is passed without using the suction member (when the liquid is passed naturally). Further, by using the suction member, the liquid passing speed can be controlled with high accuracy.
There is no restriction | limiting in particular as said suction member, According to the objective, it can select suitably, For example, a suction pump etc. are mentioned. Examples of the suction pump include a diaphragm pump and a plunger pump.

  The liquid flow can be performed using a mercury desorption solution supply member. Examples of the mercury desorption solution supply member include a syringe.

<Other processes and other means>
As said other process, a liquid passing process etc. are mentioned, for example.
Examples of the other means include liquid passing means.

-Liquid passing process and liquid passing means-
The liquid passing step is not particularly limited as long as it is a step for passing the anion exchange resin before the immobilization step, and can be appropriately selected according to the purpose.
The liquid passing means is not particularly limited as long as it is a means for passing the anion exchange resin before the immobilization step, and can be appropriately selected according to the purpose.
There is no particular limitation on the liquid used for the flow of liquid, and it can be appropriately selected depending on the purpose. However, when the sample solution contains an acid, the same kind of acid as the acid of the sample solution is used. It is preferable that the liquid is contained at a concentration because the anion exchange resin can be equilibrated.

  There is no restriction | limiting in particular as a quantity of the said liquid at the time of performing a liquid flow, Although it can select suitably according to the objective, 1 mL-10 mL are preferable and 3 mL-7 mL are more preferable.

Here, an example of the mercury analysis pretreatment method and the mercury analysis pretreatment apparatus of the present invention will be described with reference to FIGS. 1A to 1F. FIG. 1A is a schematic diagram illustrating an example of how the anion exchange resin is used. In FIG. 1A, a 5 mL syringe is used as the container 1. The filter paper 3 is placed in the lower part of the container 1, a predetermined amount of the anion exchange resin 2 is put on the filter paper 3, and the filter paper 3 is placed on the filter paper 3. I try not to leak.
FIG. 1B is a schematic diagram for explaining an example of the liquid passing process. The anion exchange resin 2 is equilibrated by passing an acid solution of the same type and the same concentration as the acid used for the sample solution through the anion exchange resin 2.
1C and 1D are schematic views for explaining an example of the immobilization process. A sample solution containing mercury and a metal other than mercury such as cadmium and iron is passed through the anion exchange resin 2. Then, mercury and some metals are fixed to the anion exchange resin 2.
FIG. 1E is a schematic diagram for explaining an example of a cleaning process. After the immobilization step, when a cleaning liquid is passed through the anion exchange resin 2, other metals than mercury are removed from the anion exchange resin 2.
FIG. 1F is a schematic diagram for explaining an example of a desorption process. When the mercury desorption solution is passed through the anion exchange resin 2 after the washing step, the mercury immobilized on the anion exchange resin 2 is desorbed from the anion exchange resin 2 and discharged out of the container 1. At this time, by using a chelating agent-containing buffer solution as a mercury desorption solution, the antibody can be used without subsequent pH adjustment or addition of a chelating agent when performing mercury analysis by immunoassay. Mercury analysis becomes possible by mixing with.

(Mercury analysis method and mercury analyzer)
The mercury analysis method of the present invention includes at least the mercury analysis pretreatment method of the present invention and a quantification step, and further includes other steps as necessary.
The mercury analyzer of the present invention includes at least the mercury analysis pretreatment apparatus of the present invention and a quantitative unit, and further includes other units as necessary.
The mercury analysis method can be suitably performed by the mercury analyzer, and the quantitative step can be suitably performed by the quantitative unit.

<Quantitative process and quantitative means>
The quantification step is not particularly limited as long as it is a step of quantifying mercury separated from a metal other than mercury by the mercury analysis pretreatment method, and can be appropriately selected according to the purpose. For example, ICP mass Analysis, ICP emission analysis, atomic absorption measurement, atomic fluorescence measurement, electrochemical measurement, nucleic acid aptamer measurement, immunoassay using antibodies, and the like. Among these, it is preferable to quantify mercury by immunoassay using an antibody (immunoassay) because it can be carried out simply, rapidly and inexpensively.
The quantification means is not particularly limited as long as it is a means for quantifying mercury separated from a metal other than mercury by the mercury analysis pretreatment method, and can be appropriately selected according to the purpose. Examples thereof include an analysis device, an ICP emission analysis device, an atomic absorption measurement device, an atomic fluorescence measurement device, an electrochemical measurement device, a measurement device using a nucleic acid aptamer, and an immunoassay device using an antibody. Among these, an immunoassay (immunoassay) apparatus using an antibody is preferable because it can be carried out simply, quickly and inexpensively.

  The antibody is not particularly limited and may be appropriately selected depending on the intended purpose. However, the hybridoma deposited under accession number FERM P-19703 in that it exhibits remarkable binding properties to the mercury-EDTA complex. Monoclonal antibodies produced from (hereinafter sometimes referred to as “Nx2C3”) are preferred.

The immunoassay is preferably measured using a measurement member (microchannel) and an optical measurement means.
The measurement member is not particularly limited and can be appropriately selected according to the purpose. The measurement sample channel (main channel) for flowing the sample to be measured and the sample flow channel to be measured are formed. A storage region (measurement site) for storing base particles (solid phase carrier) and a base material supply channel (sub-flow channel) that communicates with the storage region and allows the base particles to flow into the storage region. A measurement member used for measuring the amount of light transmitted through the sample to be measured is preferable. The measurement member has a length in a short direction of a cross section in a direction orthogonal to a flow direction of the sample to be measured in the accommodation region, so that the base particles can be arranged in a single layer in the accommodation region. It is preferable that
There is no restriction | limiting in particular as said optical measurement means, Although it can select suitably according to the objective, An absorptiometer is preferable.

  Examples of the present invention will be described below, but the present invention is not limited to these examples.

Example 1
<Reagent>
Anion exchange resin: Dowex ^{™ 1} × 2, The Dow Chemical Company (strongly basic anion exchange resin containing 2% by mass dibenylbenzene), manufactured by Wako Pure Chemical Industries, Ltd.
Dowex ^{™ 1} × 2 100-200 (particle size approximately 75 μm to 150 μm)
Dowex ^{™ 1} × 2 200-400 (particle size approximately 40 μm to 75 μm)
Ashless quantitative filter paper: B5, manufactured by Advantech Co., Ltd. Mercury standard solution: Hg 100 mg / L in 0.1M HNO ₃ , manufactured by Wako Pure Chemical Industries, Ltd. Multi-element mixed standard solution: Multielement Standard Solution W-VI, Wako Pure The content in 0.1M HNO ₃ manufactured by Yakuhin Co., Ltd. is as follows.
Al: 100 mg / L
B: 100 mg / L
Ca: 100 mg / L
Cd: 100 mg / L
Cr: 100 mg / L
Cu: 100 mg / L
Fe: 100 mg / L
Mg: 100 mg / L
Mn: 100 mg / L
Na: 100 mg / L
Pb: 100 mg / L
Zn: 100 mg / L
・ Pure water: Ultrapure water adjusted using a pure water production device (Millipore)

<Measurement equipment>
A reduction vaporization type atomic absorption device (Hg-200, manufactured by Hiranuma Sangyo Co., Ltd.) was used for analysis of mercury concentration in the measurement sample.
For analysis of other metal ions excluding mercury, an inductively coupled plasma emission spectrometer (ICP-AES, manufactured by Varian Technologies Japan Limited) was used. The analysis wavelength of each metal in the analysis by ICP-AES was as follows.
Mn: 257.610nm
Mg: 279.553 nm
Fe: 238.204 nm
Zn: 213.857 nm
Cd: 214.439 nm
Cu: 324.754 nm

(Mercury analysis pretreatment method and mercury analysis method)
Hereinafter, an example of the mercury analysis method of the present invention including the mercury analysis pretreatment method of the present invention will be specifically described in each step.
<Immobilization process>
-Preparation of anion exchange resin-filled container (measuring container)-
A 5 mL syringe (manufactured by Terumo Corporation) was used as a container. In order to prevent the anion exchange resin from dispersing in the solution to be added, add the type and amount of anion exchange resin to be described later, and use a circular ashless quantitative filter paper punched into a diameter of 13 mm for anion exchange. A structure in which the resin is sandwiched is adopted.
A specific manufacturing method is described below. First, the circular ashless quantitative filter paper was inserted into the 5 mL syringe excluding the piston and lightly pressed to fix the ashless quantitative filter paper to the bottom. Next, an anion exchange resin suspended in pure water (pure water: anion exchange resin = 1: 1 (volume ratio)) was introduced into the 5 mL syringe with a pipette so as to have a uniform thickness. Finally, a circular ashless quantitative filter paper was inserted and lightly pressed to sandwich the anion exchange resin between the filter papers to obtain a container containing the anion exchange resin.
When 5 mL of the solution was added to the completed anion exchange resin-containing container, the entire liquid flow was completed in about 10 to 15 minutes regardless of the amount of the anion exchange resin. Moreover, the liquid passing speed could be changed greatly depending on the pore diameter of the filter paper.
The anion exchange resin used in this immobilization process is shown below.
[Anion exchange resin]
Type: Dowex ^™ 1x2 100-200
Dowex ^™ 1x2 200-400
・ Amount used (when 5 mL syringe is filled): 100 μL, 500 μL

-Immobilization of mercury-
Mercury in the sample solution was immobilized on an anion exchange resin. The sample solution used is shown below.
[Sample solution]
-Hydrochloric acid solution-Hydrochloric acid concentration: 0.01 mM, 0.05 mM, 0.1 mM, 0.1 M
・ Mercury concentration: 100μg / L
-Flow rate: 5 mL

Immobilization was performed as follows. FIG. 2 is a schematic diagram for explaining the immobilization process.
Since the produced column might change its properties by drying, it was subjected to pretreatment immediately after production.
Before the immobilization step, 5 mL of hydrochloric acid solution (0.01 mM to 0.1 M) having the same concentration as the sample solution was conditioned by passing it through the anion exchange resin of the measurement container, and the syringe was filled. The anion exchange resin was equilibrated.
Subsequently, as an immobilization step, 5 mL of a sample solution having a predetermined hydrochloric acid concentration with a mercury concentration of 100 μg / L was passed through the anion exchange resin in the measurement vessel at a flow rate of 0.5 mL / min.
The passed sample solution was collected, and the amount of mercury was measured to measure the immobilization rate of mercury on the anion exchange resin.
3A to 3D show the results of the immobilization rate when the hydrochloric acid concentration of the sample solution and the type and amount of the anion exchange resin were varied.
When Dowex 1 × 2 100-200, which is a relatively large particle size anion exchange resin, was used and the amount of the anion exchange resin was 500 μL, 100% of mercury was fixed (FIG. 3A). When the amount of the anion exchange resin was 100 μL, the mercury adsorption rate was 90% to 95% at all hydrochloric acid concentrations (FIG. 3B).
On the other hand, when Dowex 1 × 2 200-400, which is a relatively small particle size anion exchange resin, was used, 100% mercury was fixed when the amount of the anion exchange resin was 500 μL (FIG. 3C). . Even when the amount of the anion exchange resin was 100 μL, 99.3% to 100% of mercury in the sample solution was fixed (FIG. 3D).

  In the following tests and processes, as a condition for immobilizing mercury without using a smaller amount of anion exchange resin, a measurement container (hereinafter, “100 μL) of anion exchange resin Dowex 1 × 2 200-400 having a relatively small particle size is used. Which may be referred to as a “standard measurement container”).

-Immobilization of contaminating metal ions-
Next, the influence of the hydrochloric acid concentration of the sample solution to be added on the immobilization of various metal ions in the sample solution on the anion exchange resin was examined. It has been shown above that immobilization of the mercury to be analyzed hardly depends on the hydrochloric acid concentration of the sample solution. Ideally, it is desirable that no immobilization of metal ions except mercury occurs. Further, even if ions other than mercury in the measurement sample are immobilized, it is considered that the influence on mercury analysis by, for example, immunoassay is small if it can be removed by washing with pure water.

As the hydrochloric acid concentration of the sample solution, 0.1 mM, 1 mM, 10 mM, 0.1 M, and 1 M were examined. To each sample solution, the multi-element mixed standard solution was added as metal ions so that the concentration of each metal ion was 100 μg / L.
After conditioning by passing 5 mL of hydrochloric acid solution (0.1 mM to 1 M) having the same concentration as the sample solution through the anion exchange resin of the standard measurement container, 5 mL of the sample solution was added to the standard measurement container. The sample solution was passed through the anion exchange resin at a flow rate of 0.5 mL / min, and the passed sample solution was collected and subjected to analysis by ICP-AES. FIG. 4 shows the immobilization ratio determined from the metal ion concentration of the sample solution having passed through each sample solution of each hydrochloric acid concentration.
When the hydrochloric acid concentration was low (0.1 mM), the cadmium ion immobilization rate was 0% and the iron ion immobilization rate was 79.9%. On the other hand, when the hydrochloric acid concentration was high (0.1 M), the iron ion immobilization rate was suppressed to 0%, but the cadmium ion immobilization rate was 26.7%. Further, when the hydrochloric acid concentration was high (1 M), the iron ion immobilization rate was suppressed to 0%, but the cadmium ion immobilization rate was 81.5%.
The immobilization rates of various metal ions differed depending on the concentration of hydrochloric acid, especially the immobilization rates of iron ions and cadmium ions. Therefore, it was confirmed that the pretreatment for mercury analysis suitable for the sample solution was possible by appropriately selecting the hydrochloric acid concentration according to the type of contaminating metal ions in the sample solution.
It was also confirmed that the hydrochloric acid concentration may be appropriately selected according to the method of mercury analysis, which is a subsequent process.

For example, when an antibody (Nx2C3 antibody) is used in an immunoassay as a mercury analysis described later, in consideration of the fact that the Nx2C3 antibody has a remarkable binding property to cadmium ions, the adsorption of cadmium ions is suppressed to 0.1 mM. It is most desirable to use a sample solution having a concentration of 10 mM hydrochloric acid.
On the other hand, the Nx2C3 antibody has a certain degree of binding to iron ions (about 1/3000 of cadmium). If the iron ions are not sufficiently removed by washing, it may become a non-negligible impurity in the immunoassay. There is sex.
Therefore, it was examined whether or not the immobilized metal ions can be removed by washing. Details will be described in “-Removal of contaminating metal ions by washing” described later.

<Washing process>
As the washing liquid in the washing step, pure water used as an eluent in an immunochromatographic simple rapid measurement kit (Cadmière) capable of detecting and quantifying Cd measurement quickly and easily was used. The flow rate of the cleaning liquid was 0.5 mL / min.

-Effect of cleaning on immobilization of mercury-
First, it was confirmed that mercury immobilized on the anion exchange resin in the standard measurement container was not desorbed by washing with pure water.
In order to examine the desorption of mercury from the anion exchange resin by washing after using the 0.1M hydrochloric acid solution with a mercury concentration of 100 μg / L as the sample solution, The water flow was repeated 5 times. And the quantity of mercury of each washing | cleaning liquid which passed was measured.
At the time of each washing, the mercury amount in the passed washing liquid was 0.3% at the maximum. The total desorption amount after 5 washes is also 0.6%. Assuming that 100% of the mercury passed through as the sample solution is fixed to the column first, 99.4% of mercury remains after washing. It was confirmed that was maintained in an immobilized state. From the above, it was shown that cleaning with pure water can be performed at least 5 times.

-Removal of contaminating metal ions by washing-
The standard measurement vessel used for the “-immobilization of contaminating metal ions” was washed 5 times with 5 mL of pure water, and the washed solution after each passage was collected for analysis by ICP-AES. The washing rate was determined from the ratio of metal ions removed by washing. The cleaning rate is shown as a percentage of each metal ion added as a sample. The result about the iron ion and cadmium ion by which the fixation was confirmed is shown to FIG. 5A and FIG. 5B. The results for zinc ions are shown in FIG. 5C.

When a sample solution having a hydrochloric acid concentration of 0.1 mM is passed, the cadmium ion immobilization rate is 1% or less as described above, while iron ions are immobilized on the anion exchange resin (79.9%). It has been shown. As a result of removing the immobilized iron ions by pure water cleaning, the cleaning rate is 4.7% to 5.8% in all of the five cleanings with pure water, and iron ions are contained in the cleaning liquid. It was confirmed that it was continuously contained (FIG. 5A).
When a sample solution having a hydrochloric acid concentration of 0.1 M was passed, the immobilization rate of cadmium ions was 26.7%. As a result of removing the immobilized cadmium ions by washing, Water washing removed 30.8% of the cadmium ions (FIG. 5B). This amount was almost the same as the total amount (26.7%) of cadmium ions in the anion exchange resin, and it was confirmed that cadmium ions could be removed by one or more washings.
That is, even if cadmium ions are immobilized on the anion exchange resin in the immobilization step, it can be easily removed by the washing step. Therefore, it was confirmed that the hydrochloric acid concentration in the immobilization step is preferably 0.1M.
In addition, when the sample solution having a hydrochloric acid concentration of 1M was passed, the immobilization ratio of zinc ions was 43.7%. As a result of removing the immobilized zinc ions by washing, Pure water washing removed 39% of the zinc ions, and the second pure water washing removed 3% of the zinc ions (FIG. 5C). This amount was equal to almost the total amount (43.7%) of zinc ions in the anion exchange resin, and it was confirmed that the zinc ions could be removed by two washes.

<Desorption step>
-Mercury desorption-
Desorption of mercury immobilized on the anion exchange resin was confirmed. The flow rate of the mercury desorption solution was 0.5 mL / min.
As the sample solution, a sample solution having a hydrochloric acid concentration of 0.1 M (mercury concentration: 100 μg / L) was used.
As the mercury desorption solution, a Tris buffer solution (pH 7.5) containing Tris buffer solution, nitric acid (0.004 mM, 0.4 mM, 40 mM), 40 mM sulfuric acid, and EDTA (ethylenediaminetetraacetic acid) was used. The EDTA, the concentration was used 10 [mu] M, EDTA-4Na of 100 [mu] M, the EDTA-4NH ₄ respectively. Moreover, about the acid used at the time of pH adjustment of a mercury desorption solution, it tested by 2 types, hydrochloric acid and nitric acid. The concentration of Tris buffer was 50 mM.

First, 5 mL of 0.1 M hydrochloric acid was passed through the standard measurement vessel for conditioning (liquid passing step), and then 5 mL of the sample solution was passed through to immobilize mercury.
Subsequently, 5 mL of various mercury desorbing solutions were passed through, the mercury desorbing solutions that were passed were collected, and the amount of mercury desorbed from the anion exchange resin was measured. The amount of mercury was expressed as a percentage of the amount of mercury added as a sample solution. FIGS. 6A to 6G show the mercury desorption rate in each desorption and the total amount of desorbed mercury finally obtained when each mercury desorption solution is used.

In the desorption step using the Tris buffer, about 93% of mercury was desorbed by the first desorption (FIG. 6A).
In the desorption step using nitric acid or sulfuric acid, mercury desorption was not confirmed when nitric acid of 0.4 mM or less was used as the mercury desorption solution, but when 40 mM nitric acid was used, In the first desorption, 55.7% mercury was desorbed, and in the second desorption, about 98.7% mercury was desorbed (FIG. 6B). In addition, even when 40 mM sulfuric acid was used, approximately the same desorption behavior as nitric acid was exhibited, and about 99.2% of mercury was desorbed by the second desorption (FIG. 6C).
In the Tris buffer containing EDTA, there was no difference in the desorption behavior depending on the type and concentration of EDTA, but a significant difference was confirmed depending on the acid used for pH adjustment. Specifically, in a mercury desorption solution (Tris buffer solution containing EDTA) adjusted for pH with hydrochloric acid, an average desorption of 20.0% to 30.0% from the first to the third desorption. In the desorption up to the fifth time, the desorption rate of mercury remained at 85.3% to 90.7% (FIGS. 6D and 6E). On the other hand, in a mercury desorption solution (Tris buffer solution containing EDTA) whose pH was adjusted with nitric acid, 75.6% to 79.75 in the first desorption regardless of the type and concentration (10 μM, 100 μM) of EDTA. 9% of mercury was desorbed, and 94.3% to 97.5% of mercury was desorbed by the second desorption (FIGS. 6F and 6G).

  Summarizing the desorption of mercury in the desorption process, when using four types of mercury desorption solutions, including Tris buffer, 40 mM nitric acid, 40 mM sulfuric acid, and Tris buffer containing EDTA and pH adjusted with nitric acid, It was possible to desorb the total amount of mercury immobilized quickly (by desorption up to the second time). Moreover, the final nitric acid concentration of the Tris buffer whose pH was adjusted with nitric acid was 36 mM to 40 mM. Furthermore, desorption of mercury could not be confirmed with nitric acid of 0.4 mM or less. From these facts, it was confirmed that the concentration of nitric acid in the mercury desorption solution is more than a certain level in desorbing mercury from the anion exchange resin.

  When using an antibody for mercury analysis, Tris buffer containing EDTA is capable of mercury analysis by mixing with antibody without adjusting pH or adding new EDTA. Especially suitable for.

  In the subsequent examination, the desorption conditions were that the mercury desorption solution was a 50 mM Tris buffer solution (pH 7.5, adjusted with nitric acid) containing 10 μM EDTA-4Na, and the number of times of liquid flow was 2 times (5 mL × 2 times). . Note that EDTA in the mercury desorption solution is considered to react rapidly with mercury at the time of desorption or after desorption to form a mercury-EDTA complex, regardless of the presence of anions such as nitric acid in the liquid.

-Desorption of contaminating metal ions-
The desorption of contaminating metal ions in the desorption process was evaluated.
The anion exchange resin of the standard measurement container in which “-immobilization of contaminating metal ions” was performed was washed with 5 mL of pure water three times, and 5 mL of mercury desorption solution (10 μM EDTA-4Na was added). The 50 mM Tris buffer solution (pH 7.5, adjusted with nitric acid) containing the solution was passed twice, and the mercury desorption solution (5 mL each) was used for analysis by ICP-AES. FIG. 7A and FIG. 7B show the desorption rates of metal ions excluding mercury by the mercury desorption solution when using sample solutions of various hydrochloric acid concentrations (0.1 mM, 0.1 M).
When a sample solution having a hydrochloric acid concentration of 0.1 mM was passed, it was shown that about 20% of iron ions that were not removed by washing were contained in the mercury desorption solution (FIG. 7A). Although the concentration of contaminating metal ions used in this example is as relatively small as 100 μg / L, in the actual environmental sample, there is a possibility of existence in the order of several mg / L (ppm). Therefore, when the hydrochloric acid concentration is 0.1 mM, the combination with other pretreatment capable of removing iron ions and the selection of an analysis method that does not affect the presence of iron ions in the mercury analysis after this pretreatment are preferable. .
On the other hand, when a sample solution having a hydrochloric acid concentration of 0.1M is passed, all the contaminating metal ions including cadmium ions are removed in the step of passing the sample solution and washing with pure water once, and the mercury desorption solution. (Fig. 7B).

From the above, it was confirmed that the preferred embodiment of the mercury analysis pretreatment method of the present invention, particularly the preferred embodiment of the mercury analysis pretreatment method of the present invention when an immunoassay is used for mercury analysis is as follows.
Hydrochloric acid concentration of sample solution: 5 mM to 0.5 M (more preferably 10 mM to 0.1 M)
Sample solution volume: 1 mL to 10 mL (more preferably 3 mL to 7 mL) x 1 Wash with pure water: 1 mL to 10 mL (more preferably 3 mL to 7 mL) x 1 to 3 times Mercury desorption solution: Contains EDTA-4Na Tris buffer (pH adjustment with nitric acid)
EDTA concentration of mercury desorption solution: 5 μM to 500 μM (more preferably 10 μM to 100 μM)
Mercury desorption solution volume: 1 mL to 10 mL (more preferably 3 mL to 7 mL) x 2 times

<Quantitative process 1>
-Quantitative means and measurement principle-
Mercury was quantified by microimmunoassay using monoclonal antibody Nx2C3 as an antibody that binds to the mercury-EDTA complex.
As a secondary antibody, a colloidal gold labeled antibody (gold colloid particle size 20 nm) was used. Carrier particles (solid phase carrier) in which a cadmium-EDTA complex was immobilized as an antigen were used. The secondary antibody and the carrier particles are described in “2.3 Gold Colloid Labeling” in the previous report (Date Yasuki, Masami Terado, Kazuhiro Sasaki, Arata Ada, Hakuo Matsumoto, Naoya Omura: Electric Power Research Laboratory Report V10030 (2011)). Preparation of antibodies and carrier particles "and the literature described in the above-mentioned report (Kazuhiro Sasaki et al., Analytical Chemistry, 56, 29-36 (2007), Kazuhiro Sasaki et al., Power Research Institute Report V08007 (2009), K. Sasaki et al., Anal. Chem., 81, 4005-4009 (2009), and J. M. Martin et al., J. Immunoassay, 11, 31-47 (1990)).
The method for preparing the carrier particles is specifically as follows. PMMA particles (GM-100SA, particle size 100 μm, manufactured by Ganz Kasei Co., Ltd.) are used as carrier particles, and BSA (bovine serum albumin) is physically adsorbed to the carrier particles, and then chelate molecules (ITCB (isotope) bonded to amino groups. Thiocyanobenzyl) -EDTA). At the time of measurement, a cadmium chloride solution was added to the suspension of carrier particles to a final concentration of 2 mM to form a metal complex, thereby obtaining a solid phase carrier to which the Nx2C3 antibody was bound.

A schematic diagram and principle of the microimmunoassay are shown in FIGS. 8A to 8C. The microimmunoassay applies the principle of a flow type immunoassay based on the binding exclusion effect to a microchannel, and enables rapid and highly accurate measurement.
As shown in FIGS. 8A and 8B, the microchannel introduces a main channel for liquid flow, a measurement site provided in the main channel, and a solid phase carrier (carrier particles) into the measurement site. And a sub-channel for introducing the carrier. At both ends of the measurement site, weirs having a width of 100 μm are provided at intervals of 50 μm so that the carrier particles do not flow out to other main flow paths.

  In the microimmunoassay, a measurement solution which is an equilibrium solution of an antigen (mercury-EDTA) and an antibody is passed through the carrier particles filled in the measurement site of the microchannel, and then captured on the carrier particles. In order to measure the obtained antibody by absorbance, a colloidal gold-labeled secondary antibody solution was passed through. The measurement solution was prepared by mixing the mercury desorption solution and the antibody solution (100-fold concentration). The concentration of the blocking reagent (N101, NOF Corporation) in the measurement solution was 1% by mass. The microimmunoassay was performed according to the method described in the previous report (Yasumoto Date, Shingo Teramon, Kazuhiro Sasaki, Arata Ada, Hakuo Matsumoto, Naoya Omura: Electric Power Research Laboratory Report V10030 (2011)). Measurement was performed using a fluidic device (microchannel) and a flow-type absorbance meter (see FIG. 8C).

-Determination of mercury-
-Preparation of sample solution and pretreatment conditions-
The measurement sample used for this determination process was prepared by the following method.
First, 0.1M hydrochloric acid to which a multi-element standard solution is added so that each metal ion is 100 μg / L is used at each concentration (0 μg / L, 0.5 μg / L, 1 μg / L, 3 μg / L, 5 μg / L). , 10 μg / L, 50 μg / L) of a sample solution containing mercury was prepared.
Then, the sample solution containing mercury at each concentration is pretreated by the mercury analysis pretreatment method of the present invention including the liquid passing step, the immobilization step, the washing step, and the desorption step under the optimum conditions described below, and then removed. The mercury desorption solution after the separation step was collected.
Container: 5 mL syringe Anion exchange resin: Dowex 1 × 2 200-400, 100 μL
Hydrochloric acid concentration of sample solution: 0.1M
Sample solution volume and number of passes: 5 mL × 1 Washing solution: pure water Wash solution volume and number of passes: 5 mL × 3 times Mercury desorption solution: 50 mM Tris buffer containing 10 μM EDTA-4Na (pH 7.5, adjustment with nitric acid) )
Mercury desorption solution volume and number of times: 5 mL x 2 times

-Microimmunoassay-
The measurement conditions by the microimmunoassay were performed according to the previous reports (Yasumoto Date, Shingo Terado, Kazuhiro Sasaki, Arata Aota, Hakuo Matsumoto, Naoya Omura: Electric Power Research Institute Report V10030 (2011)).
The measurement site in the microchannel was filled with the carrier particles on which the antigen (cadmium-EDTA) was immobilized, and then the measurement solution containing the antibody (including the mercury desorption solution) was passed therethrough. . The antibody concentration of the measurement solution was 100 pM, the feeding speed was 400 μL / min, and the feeding volume was 2 mL. Further, following the feeding of the measurement solution, 4 nM colloidal gold labeled antibody was fed as a secondary antibody solution at a feeding rate of 400 μL / min and a feeding rate of 2 mL.

The amount of unreacted antibody captured on the carrier particles was optically measured using colloidal gold colloid labeled with the antibody. FIG. 9 shows the relationship between the mercury concentration in the sample solution and the measured relative signal value.
When the measurement range was defined as a range in which the relative signal value was 10% to 90%, the measurement range was 0.5 μg / L to 7.0 μg / L. In the conventional immunoassay for heavy metals, the measurement range was two orders of magnitude or more (Kazuhiro Sasaki et al .: Analytical Chemistry (Bunsekikaku), 56, 29-36 (2007), and Yasuki Date et al .: Report of the Central Research Institute of Electric Power Industry V10030 (2011)), but in this result, the measurement range was about one digit. In addition, when the mercury concentration corresponding to the relative signal value obtained by subtracting 3 times the standard deviation from 100% when the sample containing no mercury was repeatedly measured was taken as the lower limit of detection, it was 0.6 μg / L. Moreover, it was 3.2 μg / L when 5 times the lower limit of detection was taken as the lower limit of quantification. The lower limit of detection and the lower limit of quantification are the standard value (5 μg / L) for specially controlled industrial waste (elution) specified by the Waste Management Law and mercury for wastewater discharged into public water areas specified by the Water Pollution Control Law. Satisfied.

<Quantitative process 2>
The amount of the sample solution in the immobilization process was increased, pretreatment was performed so that mercury was concentrated in the mercury desorption solution, and the quantitative process was performed using the mercury desorption solution after the pretreatment.
Specifically, in the quantification step 1, the quantification step is performed in the same manner as the quantification step 1 except that the number of passages of the sample solution is changed to the following number of passages. The relationship with the measured relative signal value was determined. The results are shown in FIG.
Sample solution amount and number of times of liquid flow: 5 mL × 10 = 50 mL
It was confirmed that the sensitivity could be increased by increasing the amount of the sample solution and concentrating mercury in the mercury desorption solution.
When the measurement range was defined as a range where the relative signal value was 10% to 90%, the measurement range was 0.1 μg / L to 1.9 μg / L. Moreover, when the mercury concentration corresponding to the relative signal value obtained by subtracting 3 times the standard deviation from 100% when the sample containing no mercury was repeatedly measured was taken as the lower limit of detection, it was 0.1 μg / L. Further, when 5 times the lower limit of detection was taken as the lower limit of quantification, it was 0.7 μg / L. The lower limit of detection and the lower limit of quantification are the standard value (5 μg / L) for specially controlled industrial waste (elution) specified by the Waste Management Law and mercury for wastewater discharged into public water areas specified by the Water Pollution Control Law. Satisfactory enough, and the lower detection limit is the world's strictest mercury standards for Japanese drinking water (0.5 μg / L, new water quality standards and inspection items by the Ministry of Health, Labor and Welfare, and inspection items (standard 51 items, Heisei 20.03. 25 announcements) and water quality management goals).

(Example 2)
The desorption process was performed using various mercury desorption solutions.
The desorption step is the same as “-Mercury desorption” in Example 1 except that the mercury desorption solution is replaced with the following mercury desorption solution in “-Mercury desorption-” in Example 1. I went there. The results are shown in FIGS. 10A to 10E.

[Mercury elimination solution]
(1) 50 mM Tris buffer (FIG. 10A)
(10 μM or 100 μmM EDTA-Na, sulfuric acid, pH 7.5)
(2) 50 mM Tris buffer (FIG. 10B)
(10 μM or 100 μmM EDTA-Na, phosphoric acid, pH 7.5)
(3) 100 mM Tris buffer (FIG. 10C)
(10 μM or 100 μmM EDTA-Na, about 80 mM HNO ₃ , pH 7.5)
(4) 500 mM Tris buffer (FIG. 10D)
(10 μM or 100 μmM EDTA-Na, about 400 mM HNO ₃ , pH 7.5)
(5) Phosphate buffered saline (PBS) (10 μM or 100 μmM EDTA-Na) (FIG. 10E)

It was confirmed that any of these mercury desorption solutions can be used.
Moreover, it was confirmed that the concentration of the Tris buffer used as the mercury desorption solution is preferably 50 mM or more, more preferably 100 mM or more.

(Example 3)
The experiment was conducted by changing the flow rate.
<Reagent>
Anion exchange resin: Dowex ^{™ 1} × 2, The Dow Chemical Company (strongly basic anion exchange resin containing 2% by mass dibenylbenzene), manufactured by Wako Pure Chemical Industries, Ltd.
Dowex ^{™ 1} × 2 200-400 (particle size approximately 40 μm to 75 μm)
Ashless quantitative filter paper: B5, manufactured by Advantech Co., Ltd. Mercury standard solution: Hg 100 mg / L in 0.1M HNO ₃ , manufactured by Wako Pure Chemical Industries, Ltd. Multi-element mixed standard solution: Multielement Standard Solution W-VI, Wako Pure The content in 0.1M HNO ₃ manufactured by Yakuhin Co., Ltd. is as follows.
Al: 100 mg / L
Ca: 100 mg / L
Cd: 100 mg / L
Cr: 100 mg / L
Cu: 100 mg / L
Fe: 100 mg / L
Mg: 100 mg / L
Mn: 100 mg / L
Pb: 100 mg / L
Zn: 100 mg / L
・ Pure water: Ultrapure water adjusted using a pure water production device (Millipore)

<Measurement equipment>
A reduction vaporization type atomic absorption device (Hg-200, manufactured by Hiranuma Sangyo Co., Ltd.) was used for analysis of mercury concentration in the measurement sample.
For analysis of other metal ions excluding mercury, an inductively coupled plasma emission spectrometer (ICP-AES, manufactured by Varian Technologies Japan Limited) was used. The analysis wavelength of each metal in the analysis by ICP-AES was as follows.
Mn: 257.610nm
Mg: 279.553 nm
Fe: 238.204 nm
Zn: 213.857 nm
Cd: 214.439 nm
Cu: 324.754 nm

<Example of pretreatment method for mercury analysis (1)>
The pretreatment method for mercury analysis was carried out by changing the flow rate using suction force.
The flow of the experimental operation in this example is shown below.
(1) Preparation of measurement container (Anion exchange resin: Dowex ^{™ 1} × 2 200-400, 100 μL)
(2) Conditioning (100 mM HCl, 5 mL)
(3) Sample solution flow using suction force (sample solution: 100 mM HCl, 10 μg / L HgCl ₂ )
(4) Washing (pure water, 15 mL)
(5) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL)

-Preparation of anion exchange resin-filled container (measuring container)-
A 5 mL syringe (manufactured by Terumo Corporation) was used as a container. In order to prevent the anion exchange resin from being dispersed in the solution to be added by placing the above-mentioned type and amount of the anion exchange resin in the container, an anion is obtained using a circular ashless quantitative filter paper punched into a diameter of 13 mm. The exchange resin is sandwiched.
A specific manufacturing method is described below. First, the circular ashless quantitative filter paper was inserted into the 5 mL syringe excluding the piston and lightly pressed to fix the ashless quantitative filter paper to the bottom. Next, an anion exchange resin suspended in pure water (pure water: anion exchange resin = 1: 1 (volume ratio)) was introduced into the 5 mL syringe with a pipette so as to have a uniform thickness. Finally, a circular ashless quantitative filter paper was inserted and lightly pressed to sandwich the anion exchange resin between the filter papers to obtain a container containing the anion exchange resin.

-Passing of sample solution using suction force and immobilization of mercury on anion exchange resin-
Using the suction force, the sample solution was passed at a higher speed.
Specifically, as shown in FIG. 11, a suction member 4 (Visprep SPE Vacuum Manifold standard, 12-port model, manufactured by SUPELCO) was attached to the tip of a container (syringe) 1 containing an anion exchange resin 2. Subsequently, 5 mL of the sample solution was placed in the syringe, and suction was performed by the suction member 4 under the pressure setting shown in Table 1, and the liquid was passed. The flow rate at that time is shown in Table 1 and FIG. It was confirmed that the flow rate can be controlled with high accuracy by controlling the pressure.

  Using the suction member, the flow rate of the sample solution was changed to fix the mercury in the sample solution to the anion exchange resin, and the immobilization rate was measured by the method described above. The results are shown in FIG. When the flow rate was 5.83 mL / min, the immobilization rate was almost 100%, and very good results were obtained.

-Desorption of mercury from anion exchange resin by liquid flow using suction force-
Mercury was desorbed from the anion exchange resin on which mercury was immobilized by changing the flow rate of the mercury desorbing solution using a suction member, and the desorption rate was measured by the method described above. The results are shown in FIG. When the flow rate was 5.83 mL / min, the desorption rate was 95%, and a very good result was obtained.

-Removal of contaminated metals-
The following experimental operation was performed, and it was confirmed whether or not contaminated metals were mixed in the mercury desorption solution when the liquid flow rate was 5.8 mL / min. The multi-element mixed standard solution was used for contamination metal contamination. The results are shown in Table 2.
The flow of operations in this experiment is shown below.
(1) Sample solution flow (flow rate 5.8 mL / min)
(Sample solution: 100 mM HCl, contaminated metal content 100 ppb)
(2) Washing (pure water, 15 mL, liquid flow rate 5.8 mL / min)
(3) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL, liquid flow rate 5.8 mL / min)
(4) ICP analysis of mercury desorption solution

  From the results of Table 2, it was confirmed that even when the flow rate of the sample solution, the wash water, and the mercury desorption solution was increased, the mercury desorption solution contained almost no contaminating metal.

  When the sample solution, the washing water, and the mercury desorption solution are passed at 5.8 mL / min, the mercury analysis pretreatment method can be performed in a short time of about 6 minutes.

<Example of mercury analysis pretreatment method (2)>
The mercury analysis pretreatment method was carried out by using a suction force to change the liquid flow rate by a method different from the above (Part 1).
The flow of the experimental operation in this example is shown below.
(1) Preparation of measurement container (Anion exchange resin: Dowex ^{™ 1} × 2 200-400, 100 μL)
(2) Conditioning (100 mM HCl, 5 mL)
(3) Sample solution flow using suction force (sample solution: 100 mM HCl, 10 μg / L HgCl ₂ )
(4) Washing (pure water, 15 mL)
(5) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL)

-Preparation of anion exchange resin-filled container (measuring container)-
The container 1 shown in FIG. 15 was used. The container 1 is composed of an upper member 1a and a lower member 1b, and when the anion exchange resin 2 is filled, the movement of the anion exchange resin 2 is restricted, and the volume of the anion exchange resin 2 is hardly changed. It has become.
The anion exchange resin 2 of the type and amount described above was placed in the container 1 and the anion exchange resin 2 was sandwiched using a circular ashless quantitative filter paper (filter paper 3) punched into a diameter of 13 mm.

-Passing of sample solution using suction force and immobilization of mercury on anion exchange resin-
Using the suction force, the sample solution was passed at a higher speed.
Specifically, as shown in FIG. 15, the syringe 5 is attached to the upper opening of the container 1 containing the anion exchange resin 2, and the suction member 4 (Visprep SPE Vacuum Manifold standard, 12-port model, SUPELCO) is attached to the lower opening. Attached). Subsequently, 5 mL of the sample solution was placed in the syringe 5 and suctioned by the suction member 4 under the pressure setting shown in Table 3, and the liquid was passed. The flow rate at that time is shown in Table 3 and FIG. It was confirmed that the liquid feeding speed could be controlled with high accuracy by controlling the pressure.

  Using the suction member, the flow rate of the sample solution was changed to fix the mercury in the sample solution to the anion exchange resin, and the immobilization rate was measured by the method described above. The results are shown in FIG. When the liquid flow rate was 2.37 mL / min and 3.44 mL / min, the immobilization rate was 90% or more, and very good results were obtained.

-Desorption of mercury from anion exchange resin by liquid flow using suction force-
Mercury was desorbed from the anion exchange resin on which mercury was immobilized by changing the flow rate of the mercury desorbing solution using a suction member, and the desorption rate was measured by the method described above. The results are shown in FIG. When the flow rate was 7.67 mL / min or less, the desorption rate was 96% or more, and a very good result was obtained.

-Removal of contaminated metals-
The following experimental operation was performed to confirm whether or not contaminated metals were mixed in the mercury desorption solution. The multi-element mixed standard solution was used for contamination metal contamination. The results are shown in Table 4.
The flow of operations in this experiment is shown below.
(1) Flow of sample solution (flow rate: 3.44 mL / min)
(Sample solution: 100 mM HCl, contaminated metal content 100 ppb)
(2) Washing (pure water, 15 mL, liquid flow rate 7.66 mL / min)
(3) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL, liquid flow rate 7.66 mL / min)
(4) ICP analysis of mercury desorption solution

  From the results in Table 4, it was confirmed that even when the flow rate of the sample solution, the washing water, and the mercury desorption solution was increased, the mercury desorption solution contained almost no contaminating metal.

<Summary of Example 3>
As in Example 3, the liquid passing speed was increased by performing the liquid passing using the suction force. As a result, the time required for the mercury analysis pretreatment method could be shortened without degrading accuracy. Specifically, the suction force is not used for passing the sample solution, the washing water, and the mercury desorbing solution, but compared with the case where only the fluid is passed by gravity (for example, in the case of Example 1). When the liquid flow rate was 5.8 mL / min using No. 1, the mercury analysis pretreatment method could be performed in 1/10 time.

Example 4
The pretreatment method for mercury analysis was performed by changing the amount of the sample solution passed through the anion exchange resin and the concentration of mercury in the sample solution.
The flow of the experimental operation in this example is shown below. In addition, the container shown in FIG. 11 was used for the container. Experiments were conducted using a suction member and setting the sample solution, the wash water, and the mercury desorption solution flow rate to 5.8 mL / min. The results are shown in FIG. Each data in FIG. 19 is an average value of three times.
(1) Preparation of measurement container (Anion exchange resin: Dowex ^{™ 1} × 2 200-400, 100 μL)
(2) Conditioning (100 mM HCl, 5 mL)
(3) Sample solution flow using suction force (sample solution: 100 mM HCl, HgCl ₂ )
(4) Washing (pure water, 15 mL)
(5) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL)

  When the mercury concentration in the sample solution is in the range of 1 μg / L to 10 μg / L and the amount of the sample solution is in the range of 5 mL to 100 mL, the mercury is immobilized on the anion exchange resin and the mercury immobilized on the anion exchange resin There was no significant change in desorption to the mercury desorption solution. That is, even if the amount of the sample solution was increased, mercury desorption into the mercury desorption solution (10 mL) occurred at almost the same rate, so it was confirmed that mercury could be concentrated.

-Removal of contaminated metals-
The following experimental operation was performed to confirm that no contaminating metal was mixed in the mercury desorption solution (removal rate) when the amount of the sample solution was changed. The results are shown in Table 5.
The flow of operations in this experiment is shown below.
(1) Sample solution flow (flow rate 5.8 mL / min)
(Sample solution: 100 mM HCl, contaminated metal content 100 ppb)
(2) Washing (pure water, 15 mL, liquid flow rate 5.8 mL / min)
(3) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL, liquid flow rate 5.8 mL / min)
(4) ICP analysis of mercury desorption solution

  From the results in Table 5, it was confirmed that even when the amount of the sample solution was increased, the mercury desorption solution contained almost no contaminating metal.

<Summary of Example 4>
Even when the amount of the sample solution is increased, mercury desorption to the mercury desorption solution (10 mL) occurs at almost the same rate, and the mercury desorption solution contains almost no contaminating metals. It was confirmed that concentration was possible.

(Example 5)
The mercury analysis pretreatment method was carried out by changing the amount of the sample solution, and the mercury quantification of the mercury desorption solution obtained as a result was determined using monoclonal antibody Nx2C3 as an antibody that binds to the mercury-EDTA complex. Performed by the microimmunoassay used. The details of the microimmunoassay method are the same as those in the quantitative step 1 of Example 1.

The flow of the experimental operation in this example is shown below. Note that the container shown in FIG. 11 was used. Experiments were conducted using a suction member and setting the sample solution, the wash water, and the mercury desorption solution flow rate to 5.8 mL / min.
(1) Preparation of measurement container (Anion exchange resin: Dowex ^{™ 1} × 2 200-400, 100 μL)
(2) Conditioning (100 mM HCl, 5 mL)
(3) Sample solution flow using suction force (sample solution: 100 mM HCl, HgCl ₂ (arbitrary amount), 5 mL to 100 mL)
(4) Washing (pure water, 15 mL)
(5) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL)

Various concentrations of mercury (0.01 μg / L, 0.04 μg / L, 0.1 μg / L, 0.4 μg / L, 1 μg / L, 4 μg / L, 10 μg / L) were used to create a calibration curve. In addition, 100 mM hydrochloric acid to which various impurities were added so as to be 100 μg / L was prepared as a sample solution. Subsequently, mercury analysis pretreatment was performed using different amounts (5 mL, 10 mL, 50 mL, 100 mL) of sample solutions, and a calibration curve was prepared by subsequent immunoassay.
The results are shown in FIG. Under each experimental condition (sample solution amount), a sigmoidal calibration curve was obtained, indicating that there was no inhibition of the antigen-antibody reaction by contaminating metals.
Table 6 shows the lower limit of detection under each condition. The lower limit of detection (0.12 μg / L) when using 50 mL of the sample solution and the lower limit of detection (0.04 μg / L) when using 100 mL of the sample solution are the mercury standards for drinking water in Japan. The value (0.5 μg / L, Ministry of Health, Labor and Welfare ordinance new water quality standard and inspection items (standard item 51 / 20.03.25 issue) / water quality management target) was extremely low.

  The lower limit of detection in Table 6 was calculated by the 3σ method (σ: blank standard deviation).

(Example 5)
<Analysis of standard environmental samples>
A mercury analysis pretreatment method was implemented for standard environmental samples, followed by quantitative analysis of mercury using an immunoassay. In addition, since the amount of mercury contained in each standard sample is known, the volume of the sample solution used for mercury analysis pretreatment can be quantified with sufficient accuracy with reference to a calibration curve (FIG. 20). Selected to fall within range (measurement range IC80-IC20).

-Analysis of river water samples-
Mercury was added to an inorganic analysis river water certified substance (JSAC0301-2), which is a standard sample, to a concentration of 0.5 μg / L (environmental reference value) to prepare a 50 mL sample solution.
The mercury analysis of the present invention was performed using the sample solution prepared above. The flow of the experimental operation in this example is shown below. In addition, the container shown in FIG. 11 was used for the container. Experiments were conducted using a suction member and setting the sample solution, the wash water, and the mercury desorption solution flow rate to 5.8 mL / min.
(1) Preparation of measurement container (Anion exchange resin: Dowex ^{™ 1} × 2 200-400, 100 μL)
(2) Conditioning (100 mM HCl, 5 mL)
(3) Sample solution flow using suction force (sample solution: 50 mL)
(4) Washing (pure water, 15 mL)
(5) Desorption (50 mM Tris-HNO ₃ (pH 7.5) +10 μM EDTA, 10 mL)
(6) Quantification by immunoassay (the same method as quantification step 1 of Example 1)

  The quantification by immunoassay was performed using a calibration curve, and the recovery rate was calculated from the average value of five analysis values. The results are shown in Table 7. FIG. 21 shows the relationship between the measurement result and the calibration curve. The mercury concentration calculated from the average value of the five measurements was 0.47 μg / L. Since the recovery rate of the analysis for the amount of added mercury was as high as 94.3 ± 9.4%, the mercury analysis method of the present invention is a river water sample near the environmental standard value (0.5 μg / L). It was shown to be useful for mercury analysis. In addition, it is considered possible to analyze tap water having the same composition.

-Analysis of soil samples-
When the test solution prepared by the method according to “Ministry of the Environment Notification No. 19, Determination of Measurement Method for Soil Content Survey” (March 06, 2003) was subjected to instrumental analysis, the mercury content was (0.61 ± 0.08 mg / kg). This value was in good agreement with the certified value (0.6 ± 0.1 mg / kg) of the standard substance (JSAC0402). From this, it was shown that the prepared test solution is appropriate.

Mercury analysis was performed under the same conditions as in the above “Analysis of river water sample” except that 10 mL of the test solution prepared as the sample solution used for mercury analysis was used.
Table 8 shows the results. FIG. 22 shows the relationship between the measurement result and the calibration curve. The quantitative value of mercury in the measurement solution of the immunoassay calculated from the average value of the five measurements was 1.65 μg / L. Here, as a result of calculating the mercury content of the soil measured from the solid-liquid ratio at the time of preparing the test solution, the concentration ratio at the time of the mercury analysis pretreatment, the mercury content of the soil sample was 0.55 ± 0.12 mg. / Kg was calculated. A comparatively good recovery of the analysis (90.2 ± 6.8%) was obtained compared to the analytical value (0.61 mg / kg) measured by the instrument previously measured.

-Analysis of coal ash sample (simulated dissolution test)-
When the test solution prepared by the method according to “Environment Agency Notification No.13, Test Method for Metals in Industrial Waste” (February 17, 1973) was subjected to instrumental analysis, the test solution was mercury. (Below the lower limit of detection) Mercury was added so that the amount would be 5 μg / L (elution amount base value in the management-type landfill of the waste treatment method), and the quantitativeness of mercury contained in the test solution was increased. evaluated.
Mercury analysis was performed under the same conditions as in the above “Analysis of River Water Sample” except that 5 mL of the prepared test solution was used as an appropriate amount of sample solution for pretreatment for mercury analysis.
The results are shown in Table 9. FIG. 23 shows the relationship between the measurement result and the calibration curve. The mercury concentration calculated from the average value of the five measurements was 5.2 μg / L. Here, the recovery rate of the analysis for the added mercury amount (5 μmg / L) is 104.8 ± 5.8%, and the elution amount base value (5 μg / L, in the test solution) in the management type landfill of the waste treatment method ) Quantitative analysis was shown in the vicinity.

  The mercury analysis pretreatment method and the mercury analysis pretreatment apparatus of the present invention can separate the mercury and the metal other than mercury from a liquid containing mercury and a metal other than mercury. And as a pretreatment device.

Aspects of the present invention are as follows, for example.
<1> A mercury analysis pretreatment method for separating the mercury and the metal other than mercury from a sample solution containing mercury and a metal other than mercury,
An immobilization step of passing the sample solution through an anion exchange resin and immobilizing at least the mercury on the anion exchange resin;
Subsequently, the anion exchange resin is washed with washing water, and a washing step for removing metals other than mercury from the anion exchange resin,
Subsequently, a mercury desorption solution for desorbing the mercury from the anion exchange resin is passed through the anion exchange resin, and a desorption step of desorbing the mercury from the anion exchange resin is included. It is a characteristic mercury analysis pretreatment method.
<2> The mercury analysis pretreatment method according to <1>, wherein the mercury desorption solution contains a buffer.
<3> The mercury analysis pretreatment method according to <2>, wherein the mercury desorption solution further contains a chelating agent that chelates mercury to form a mercury complex.
<4> The mercury analysis pretreatment method according to any one of <1> to <3>, wherein the mercury desorption solution contains at least one of nitric acid, sulfuric acid, and phosphoric acid.
<5> The mercury analysis pretreatment method according to any one of <1> to <4>, wherein the mercury desorption solution contains at least one of nitric acid and sulfuric acid.
<6> At least one of the sample solution flow in the immobilization step, the wash water flow to the anion exchange resin in the wash step, and the mercury desorption solution flow in the desorption step uses the suction member. The mercury analysis pretreatment method according to any one of <1> to <5>, which is performed.
<7> A mercury analysis pretreatment apparatus for separating mercury and the metal other than mercury from a sample solution containing mercury and a metal other than mercury,
Immobilization means for passing the sample solution through an anion exchange resin and immobilizing at least the mercury on the anion exchange resin;
Washing means for washing the anion exchange resin with washing water, and removing metals other than mercury from the anion exchange resin;
A mercury desorption solution for desorbing the mercury from the anion exchange resin is passed through the anion exchange resin, and desorption means for desorbing the mercury from the anion exchange resin. Mercury analysis pretreatment equipment.
<8> A mercury desorption solution characterized by being used to desorb mercury from an anion exchange resin containing a buffer and having mercury immobilized thereon.
<9> The mercury desorption solution according to <8>, further including a chelating agent that chelates mercury to form a mercury complex.
<10> The mercury desorption solution according to <9>, wherein the chelating agent is ethylenediaminetetraacetic acid.
<11> The mercury desorbing solution according to any one of <8> to <10>, wherein the buffer is trishydroxymethylaminomethane.
<12> The mercury analysis pretreatment method according to any one of <1> to <6>,
And a quantitative step of quantifying mercury separated from a metal other than mercury by the mercury analysis pretreatment method.
<13> The mercury analysis method according to <12>, wherein the quantification step quantifies mercury by an immunoassay using an antibody.
<14> The mercury analysis method according to <13>, wherein the antibody is a monoclonal antibody produced from a hybridoma deposited under accession number FERM P-19703.
<15> The mercury analysis pretreatment device according to <7>,
The mercury analyzer has a quantitative means for quantitatively determining mercury separated from a metal other than mercury by the mercury analysis pretreatment device.

1 Container 2 Anion Exchange Resin 3 Filter Paper 4 Suction Member 5 Syringe

Claims (11)

A mercury analysis pretreatment method for separating the mercury and the metal other than mercury from a sample solution containing mercury and a metal other than mercury,
An immobilization step of passing the sample solution through an anion exchange resin and immobilizing at least the mercury on the anion exchange resin;
Subsequently, the anion exchange resin is washed with washing water, and a washing step for removing metals other than mercury from the anion exchange resin,
Then, it passed through the anion mercury desorbing the mercury exchange resin desorption solution to the anion exchange resin, saw including a desorption step of the mercury is desorbed from the anion exchange resin,
The mercury desorption pretreatment method, wherein the mercury desorption solution contains a buffer and at least one of nitric acid, sulfuric acid, and phosphoric acid, and has a pH of 1.0 to 8.5 . The mercury analysis pretreatment method according to claim 1, wherein the mercury desorption solution contains a buffer and at least one of nitric acid and sulfuric acid, and has a pH of 1.0 to 8.5. The mercury analysis pretreatment method according to any one of claims 1 to 2, wherein the mercury desorption solution further contains a chelating agent that chelates the mercury to form a mercury complex. The liquid passage of the sample solution in the fixing step, wherein the wash water passing liquid to the anion exchange resin in the washing step, and the at least one of liquid passing the mercury desorption solution in the desorption step the suction The mercury analysis pretreatment method according to claim 1, wherein the mercury analysis is performed using a member. A mercury analysis pretreatment apparatus for separating mercury and the metal other than mercury from a sample solution containing mercury and a metal other than mercury,
Immobilization means for passing the sample solution through an anion exchange resin and immobilizing at least the mercury on the anion exchange resin;
Washing means for washing the anion exchange resin with washing water, and removing metals other than mercury from the anion exchange resin;
The mercury desorption solution to desorb the mercury from the anion-exchange resin was passed through the anion exchange resin, the mercury possess a desorbing means for desorbing from the anion exchange resin,
The mercury desorption pretreatment apparatus, wherein the mercury desorption solution contains a buffer and at least one of nitric acid, sulfuric acid, and phosphoric acid, and has a pH of 1.0 to 8.5 . And buffering agents, nitric acid, containing at least any of sulfuric acid and phosphate, pH is 1.0 to 8.5, using the anion exchange resin mercury is immobilized in order to the mercury desorbed A mercury desorption solution characterized in that The mercury desorption solution according to claim 6, wherein the buffer is trishydroxymethylaminomethane. A mercury analysis pretreatment method according to any one of claims 1 to 4,
Mercury analysis method characterized by comprising a quantitative quantifying the mercury is separated from the metal other than the mercury by the mercury analysis pretreatment method. The quantification step, mercury analysis method according to claim 8 for quantifying the mercury by immunoassay using an antibody. The mercury analysis method according to claim 9, wherein the antibody is a monoclonal antibody produced from a hybridoma deposited under accession number FERM P-19703. The mercury analysis pretreatment device according to claim 5;
Mercury analyzer and having a dosing means for quantifying the mercury is separated from the metal other than the mercury by the mercury analyzer pretreatment device.