JP2003194798A - Method for determining a small amount of environmentally polluting substance and kit for determination used therefor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a method for easily and accurately determining an extremely small amount of components, especially heavy metal ions, causing environmental pollution. <P>SOLUTION: A heat-reversible heat-sensitive high-molecular compound of a fine-fiber-like solid which is hydrophilic at low temperatures and hydrophobic at high temperatures is added to a supernatant liquid of the extremely small amount of pollutants, dissolved, and heated to a transition temperature or more to form solid matter and separate a liquid part. The solid heat-sensitive high- molecular compound encloses a hydrophobic coloring reagent which comes into reaction with the extremely small amount of pollutants and is colored. A solid part made of the heat-sensitive high-molecular compound mixed at temperatures lower than the transition temperature, heated to the transition temperatures or higher, enclosing a colored compound generated by the reaction between the extremely small amount of pollutants and the coloring reagent, and deposited is separated from the liquid part, dissolved in water or a solvent to measure its absorbance, or calorimetrically determined with a standard item. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel method for quantifying a trace amount of environmental pollutants and a quantification kit used therefor.

[0002]

2. Description of the Related Art In recent years, from the viewpoint of preventing environmental pollution, it has become necessary to quantify very small amounts of components in the air, water or soil. In addition, there is an increasing demand for a method of accurately quantifying. As one of the analysis methods for such trace components, there is a colorimetric determination method for heavy metal ions. This is a method of reacting an analyte with a color-developing reagent, pre-concentrating the obtained reaction product, and then performing spectroscopic analysis.
It is used in a wide range of fields.

However, in this method, preconcentration is an indispensable step for enhancing the analytical sensitivity, and it is carried out by means such as evaporative concentration, solvent extraction, coagulation, dialysis, etc., but the operation is complicated in all cases. It takes a long time,
There are problems such as high cost, unstable samples, and the need to use an organic solvent that may cause carcinogenesis, and the use range is unavoidable.

As a measure against such a problem, a coloring reagent having a large molecular extinction coefficient which reacts specifically with the substance to be analyzed ["Fine Chemical", Vol. 20, No. 6, No. 5].
~ 12 (1991)] and a method of concentrating a reaction product of an analyte and a color reagent with a membrane filter ["Analyst", Vol. 112, pp. 1257-1260 (1987)] and the like. Although proposed, it is not always satisfactory for practical use.

On the other hand, in the field of environmental analysis, in particular, it is practiced to collect a sample at a site and bring the sample back to a place equipped with an analytical instrument for analysis. Although this method has the advantage of high analysis accuracy, it has the disadvantages that it takes a long time to obtain the analysis results and is expensive, so recently, on-site analysis where results can be obtained on the spot is performed. Is desired.

By the way, in this on-site analysis, no special device or instrument is required for separation and detection of the sample, and the result can be obtained quickly and with high sensitivity by simple operation.
Moreover, the cost is required to be sufficiently low.

[0007]

SUMMARY OF THE INVENTION The present invention has been made for the purpose of providing a method for easily and accurately quantifying a trace amount of components, particularly heavy metal ions, which causes environmental pollution, and a quantification kit used therefor. It is a thing.

[0008]

[Means for Solving the Problems] As a result of intensive studies to develop a method capable of easily quantitatively analyzing a trace amount of pollutants, the present inventors have found that it is hydrophilic at low temperatures and hydrophobic at high temperatures. By utilizing the characteristics of the thermoreversible thermosensitive polymer, it was found that a trace contaminant can be easily and accurately quantified, and the present invention has been completed based on this finding.

That is, according to the present invention, (a) a step of mixing an environmentally-collected sample containing trace contaminants with water, stirring them well and then leaving them to stand, and collecting the supernatant liquid and mixing with the pretreatment liquid. (B) A thermoreversible thermosensitive polymer compound of fine fibrous solid which is maintained at an arbitrary temperature of the above mixture and becomes hydrophilic at a low temperature having a transition temperature higher than that temperature and hydrophobic at a high temperature is added. A step of dissolving, (c) a step of raising the temperature of the solution above the transition temperature of the thermosensitive polymer compound to form a solid, and then separating a liquid part from the solid, (d) the liquid part A thermoreversible thermosensitive polymer compound which is hydrophilic at a low temperature and becomes hydrophobic at a high temperature in a solid state in which a hydrophobic color-forming reagent that reacts with the above-mentioned trace contaminants is included, and lower than its transition temperature. Mix at temperature to form a solution Step (e) The temperature of this solution is raised above the transition temperature, and a solid portion composed of the above heat-sensitive polymer compound is deposited by clathrating and depositing a coloring compound produced by the reaction of a trace contaminant and the above-mentioned coloring reagent. A step of separating from the liquid portion, (f) a step of dissolving the solid portion obtained in (e) in water or a solvent at a temperature lower than the transition temperature of the thermosensitive polymer compound to form a solution, and (g) Suitable for use in the method for quantifying environmental pollutants, which comprises the step of measuring the absorbance of a solution or colorimetrically determining with a standard product to determine the content of trace pollutants. , (A) a test tube containing a required amount of pretreatment liquid and having a marked line for adding a sample, (B) a thermoreversible thermosensitive polymer that is hydrophilic at a required amount of low temperature and hydrophobic at an elevated temperature A test tube containing the compound, (C After treatment deposited hydrophilic tube wall of (B) - a test tube for receiving the separated liquid portion from the hydrophobic thermally reversible polymeric compound solid, (D)
A necessary amount of a thermoreversible polymer compound, which is hydrophilic at low temperature and hydrophobic at high temperature, which contains a hydrophobic coloring reagent that develops color by reacting with a trace contaminant to be quantified, and (C) A test tube with a marked line for adding a liquid portion therein, (E) a small container containing a solvent for dissolving solid matter attached to the tube wall of (D) after the above treatment, and (F) It is intended to provide a kit for quantifying an environmental pollutant trace substance, which is a combination of a colorimeter printed with a colorimetric band corresponding to the concentration of the trace pollutant determined by a standard reagent.

[0010]

BEST MODE FOR CARRYING OUT THE INVENTION A thermoreversible thermosensitive polymer compound which is hydrophilic at a low temperature and hydrophobic at a high temperature used in the present invention is a hydrate of a polymer produced at a predetermined temperature, that is, a low temperature below a transition temperature. (Oxonium hydroxide) has a physical property that macromolecules aggregate and precipitate by dehydration at a temperature higher than the above temperature,
Below the transition temperature it is hydrophilic and soluble in water, but above the transition temperature it is hydrophobic and precipitates.

In the present invention, the transition temperature is 0 to 90.
A hydrophilic-hydrophobic thermoreversible thermosensitive polymer compound having a temperature of ℃, preferably 10 to 60 ° C. is used, but it has a transition temperature that shows hydrophilicity especially at ambient temperature and shows hydrophobicity when heated. Those are preferable. This transition temperature can be changed depending on the types of monomers constituting the polymer compound and the content ratio of the comonomer, but it can also be changed by adjusting the salt concentration and pH of the aqueous solution. it can.

The thermoreversible thermosensitive polymer compounds which are hydrophilic at low temperatures and hydrophobic at high temperatures are known substances, and examples thereof include N-ethyl acrylamide, Nn-propyl acrylamide and N-ethyl acrylamide. Isopropylacrylamide, N-cyclopropylacrylamide, N, N-ethylmethylacrylamide, N, N-diethylacrylamide, N-acryloylpyrrolidine,
Polymers or copolymers of at least one monomer selected from N-acryloylpiperidine, N-acryloylmorpholine and corresponding methacrylamide derivatives, or these monomers and hydrophilic monomers, for example N-vinylpyrrolidone, vinylpyridine, acrylamide, methacrylamide, N-methylacrylamide, hydroxyethyl methacrylate, hydroxyethyl acrylate, hydroxymethyl acrylate, unsaturated carboxylic acids such as acrylic acid and methacrylic acid, and salts thereof, vinyl sulfonic acid Unsaturated sulfonic acid such as styrene sulfonic acid, N, N-dimethylaminoethyl methacrylate, N, N-diethylaminoethyl methacrylate,
N, N-dimethylaminopropyl acrylamide, etc.
Alternatively, hydrophobic monomers such as acrylic or methacrylic acid esters such as ethyl acrylate, methyl methacrylate, glycidyl methacrylate, N
-N-butylacrylamide, N-alkyl-substituted acrylamide such as Nn-butylmethacrylate or methacrylamide, vinyl chloride, acrylonitrile, styrene, vinyl acetate and the like at least one monomer selected from Examples thereof include polymers.

In general, copolymerization with a hydrophilic monomer causes a higher transition temperature than a homopolymer, and copolymerization with a hydrophobic monomer causes a lower transition temperature than a homopolymer. Utilizing this, hydrophilicity with desired transition temperature-
A hydrophobic thermoreversible thermosensitive polymer compound can be formed.

By the way, generally, trace substances that cause environmental pollution are usually contained in a huge matrix such as soil or drainage. The substance needs to be separated. Until now, this separation was carried out by derivatizing a trace substance into a hydrophobic substance, for example, a chelate compound, and then extracting it from a huge matrix using a hydrophobic organic solvent such as chloroform or ether. The method requires a large amount of organic solvent and is not always satisfactory in terms of extraction efficiency.

However, an aqueous solution containing the thermoreversible thermosensitive polymer compound in a concentration of 0.01 to 0.5% by mass is a solvent having a polarity close to that of a lower alcohol and having a hydrophobicity at a certain concentration or more. In addition, it has the property of being dehydrated by temperature stimulation to form a solid phase and separated from the aqueous phase. When a hydrophobic matrix coexists, it is taken into the solid phase of the thermoreversible thermosensitive polymer compound and removed from the aqueous phase. At this time, the partition coefficient between the aqueous phase and the solid phase is determined by the hydrophobicity of the matrix substance. The higher the quality, the larger.

Therefore, when the trace substance to be quantified is kept hydrophilic and the above thermoreversible thermosensitive polymer compound is added at a temperature below its transition temperature and then heated above that transition temperature, the matrix substance Can be collected and separated. Then, this separation method does not require any equipment, simply by collecting the supernatant to be quantified by tilting, it can be performed easily and in a short time,
It is suitable as a processing method in on-site analysis.

In the method of the present invention, by utilizing the property of the thermoreversible thermosensitive polymer compound which becomes hydrophilic at a low temperature and hydrophobic at a high temperature, a trace amount of contaminants to be quantified from a sample taken from the environment is used. Separate the material. The trace contaminant may be an inorganic substance such as heavy metal ions or an organic substance such as formaldehyde.

That is, first, a predetermined amount of a sample is taken from the environment, mixed with water to transfer a trace substance in the sample into water, and collected as a supernatant to obtain a predetermined standard, for example, in the case of soil. "Environmental standards for soil pollution"
(A step) of preparing a sample solution according to the preparation method shown in (August 23, 1991, Environmental Agency Notification No. 46).

Next, a predetermined amount of this sample solution was kept at a temperature lower than the transition temperature of the thermoreversible thermosensitive polymer compound which becomes hydrophilic at the low temperature and hydrophobic at the high temperature, and the fine fibrous solid Add a thermoreversible thermosensitive polymer,
Mix well and dissolve [(b) step]. In this process the mixture forms a single aqueous phase.

Next, this mixture is heated to a temperature above the transition temperature of the thermoreversible thermosensitive polymer. If a solid substance is produced at this stage and a matrix substance is present, it is incorporated into the solid phase, so if only the liquid portion is separated from the solid substance by tilting or the like, a test liquid in which the matrix substance is removed is obtained. Obtained [(c) step].

Next, the trace substance detection material prepared in advance in this test solution is more than the transition temperature of the thermoreversible thermosensitive polymer compound contained in the material, which is hydrophilic at low temperature and hydrophobic at high temperature. Add at low temperature and dissolve [step (d)].

This trace substance detection material is prepared by including a hydrophobic chelate-forming color reagent in a thermoreversible thermosensitive polymer compound which is hydrophilic at low temperature and hydrophobic at high temperature. As the hydrophobic chelate-forming color-forming reagent to be clathrated, any one can be arbitrarily selected from those which have been used as a color-forming reagent for forming a chelate compound with a predetermined trace contaminant in photoanalysis.

Examples of such a coloring reagent include dithizone which forms a red complex with cadmium Cd (II) for the detection of heavy metal ions, 2-nitroso-1-naphthol which forms a red complex with cobalt Co (II), Chrome Cr
Diphenylcarbazide which forms a red-violet complex with (IV), thiothenoyltrifluoroacetone which forms a yellow complex with mercury Hg (I) and Hg (II), and lead Pb (I
I) and dizone which form a red complex.

In addition, as a color-forming reagent for organic environmental pollutants such as formaldehyde, a combination of sulfite-decolorized dye fuchsin, N-benzenesulfonylhydroxylamine and iron chloride, 4-amino-3-hydrazino-5 is used.
-Mercapto-1,2,4-triazole, chromotropic acid, etc.

The material for detecting trace contaminants used in the method of the present invention is prepared by dissolving the above-mentioned hydrophobic chelate-forming color-developing reagent in a water-miscible organic solvent such as acetone, dimethylformamide or dimethylsulfoxide. It can be produced by mixing the above-mentioned aqueous solution of the hydrophilic-hydrophobic thermoreversible thermosensitive polymer compound at a transition temperature or lower and lyophilizing the mixture.

At this time, the concentration of the color forming reagent in the organic solvent is 0.5 to 1.0% by mass, and
The concentration of the thermosensitive polymer compound in the aqueous solution is 0.2.
Within the range of ˜15% by mass is used. Freeze-drying is, for example, 0.5-10 at a temperature of -20 ° C to -50 ° C.
It can be performed under reduced pressure of 0 Pa. In this way
A cotton-like trace contaminant detection material in which the coloring reagent is uniformly included in the thermosensitive polymer compound is obtained.

Up to now, the organic coloring reagent has been used as a solution dissolved in an organic solvent, but the stability of the solution is low and it often deteriorates in a short time. However,
Since the above-mentioned material for detecting trace contaminants is solid, it is stable, and can sufficiently support field analysis under severe measurement conditions. Moreover, the coexistence with the polymer compound allows the chelate-forming color-forming reagent, which is usually poorly soluble in water, to be easily mixed with water to allow the color reaction to proceed rapidly.

In the method of the present invention, the test solution containing the trace contaminant detection material is heated to a temperature above the transition temperature of the thermosensitive polymer compound. By this treatment, the hydrophobic coloring compound produced by the reaction between the trace contaminant and the coloring reagent is deposited in a solid state while being included in the thermosensitive polymer compound,
Since it adheres to the vessel wall, the solid part is left as it is, and only the liquid part is removed by tilting [(e) step].

Next, the solid portion attached to the vessel wall is dissolved using a predetermined amount of water or a solvent having a temperature lower than the transition temperature of the heat-sensitive polymer compound using the heat-sensitive polymer compound to obtain a colored solution [(f) step ].

The amount of the trace contaminant contained in the colored solution thus obtained is determined by measuring its absorbance with a photometer or by colorimetrically quantifying it with a standard product [( G) process]. In this way, trace contaminants can be easily and quickly quantified on-site from a sample collected from the surrounding environment.

It is advantageous that the quantification in the above step (g) is performed by comparing the color with a standard color table prepared in advance based on a standard of known concentration. It is also possible to compare the absorbance at the absorption wavelength with that of a standard of known concentration.

The operation at low temperature in the method of the present invention is 5 to
Since it is preferable to carry out the operation in the range of 10 ° C. and the operation at high temperature in the range of 20 to 45 ° C., it is advantageous to select and use a thermosensitive polymer compound having a transition temperature in this range.

The method of the present invention is preferably carried out using a quantification kit in which tools are combined so that the operation of each step can be carried out smoothly. FIG. 1 is a perspective view showing an example of such a quantitative kit. A method for quantifying hexavalent chromium using a quantification kit will be described with reference to this figure. This kit includes a case 1, three round-bottom test tubes 2, 4 and 5 and one flat-bottom test tube 6, a small bottle 7 containing a solvent, a dropper 8 and a colorimeter 9 (colorimetric band 10 as an accessory). Including)). Then, a sample to be quantified is first stored in advance in a pretreatment liquid, for example, perchloric acid mixed liquid 3, and injected into a test tube 2 marked with a mark indicating the required amount of the sample to the position of the marked line, Mix well with pretreatment solution.

Next, the test tube 4 containing the thermoreversible thermosensitive polymer compound which becomes hydrophilic at low temperature and hydrophobic at high temperature is kept at a temperature lower than the transition temperature of the thermoreversible thermosensitive polymer compound, A sample is injected from the test tube 2 into it. Next, the test tube 4 is heated to a temperature not lower than the transition temperature of the thermoreversible polymer compound and shaken vigorously to adhere the deposited solid to the tube wall, and only the liquid portion is transferred to the test tube 5 by tilting.

Next, a flat-bottom test was conducted in which a thermoreversible polymer compound, which included a hydrophobic color-forming reagent such as diphenylcarbazide, was hydrophilic at a low temperature and hydrophobic at a high temperature, and a marked line was drawn at a required position. After the liquid in the test tube 5 was cooled to a low temperature and added to the position of the marked line of the tube 6 to dissolve the contents, the tube was heated above the transition temperature of the thermoreversible polymer compound and shaken vigorously. After the produced solid is attached to the wall of the tube, the liquid portion is discarded by tilting, leaving only the solid portion. Next, a small amount of solvent is poured from the vial 7 into the test tube 6 with a dropper 8 to completely dissolve the solid mass to obtain a coloring liquid.

The flat-bottomed test tube 6 containing the coloring liquid thus prepared is inserted into a predetermined hole of the colorimeter and compared with the color of the colorimetric band through the peep hole to find the point where both match. By reading the corresponding value, the amount of chromium can be determined. Of the test tubes, the test tube 6 to be inserted into the colorimeter has a flat bottom when compared with the color of the colorimetric band, but the other test tubes do not need to have a flat bottom.

[0037]

According to the method of the present invention, a trace amount of environmental pollutants can be rapidly quantified on-site by a simple operation, especially by using a predetermined quantification kit.

[0038]

The present invention will be described in more detail by way of examples, which should not be construed as limiting the invention thereto.

Reference Example 1 Poly (N-isopropylacrylamide) having a transition temperature of about 30 ° C. and a mass average molecular weight of about 50,000 (hereinafter referred to as PNIPA).
(Abbreviated as Am) 4 g, diphenylcarbazide 0.02 g
Was added to the solution dissolved in 5 ml of acetone and dissolved.
Diluted by adding water at ℃ until the total amount became 100 ml.
Then, this solution is placed at a temperature of -40 ° C. and a pressure of 10 Pa for 24 hours.
PNIPAAm (hereinafter DPC-PNIP) containing cotton-like diphenylcarbazide by freeze-drying for a long time
Abbreviated as AAm).

Reference Example 2 0.2 g of the same PNIPAAm as used in Example 1,
0.001 g of dithizone was added to and dissolved in a solution of 5 ml of acetone (or ethyl alcohol), and water at 5 ° C. was added to the total amount of 100 ml for dilution. Then, this solution was freeze-dried under the same conditions as in Reference Example 1 to include dithizone-encapsulated PNIPAAm (hereinafter referred to as D
TZ-PNIPAAm) was obtained.

Example 1 0.971 g of potassium chromate was weighed and the total amount was 50 ml.
Water was added to the solution until it became dissolved, and a chromic acid standard stock solution having a concentration of 0.1 M was prepared. Next, this chromic acid standard stock solution was diluted with water to prepare 10 ml of each chromium standard solution having a chromium content of 5.2 to 156 ppb. 10 ml of each of these chromium standard solutions was placed in a test tube with a lid, and 3 ml of a perchloric acid mixed solution (8M sodium perchlorate + 0.0025M perchloric acid) was added thereto and mixed, and then obtained at 5 ° C. in Reference Example 1. DPC-PNIPAAm was added and shaken to dissolve. Further, hold at this temperature for 5 minutes, Cr-D
After completing the PC complex formation reaction, the mixture was heated to 45 ° C. for 1 minute and immediately vigorously shaken for 10 seconds to adhere the produced solid to the test tube wall. Then, the liquid portion was removed by decantation, and 0.5 ml of acetone was added to dissolve the solid portion to obtain a red solution. The results of measuring the absorbance at a wavelength of 540 nm for this solution are shown in FIG. The black circles in the figure are the results when the heavy metal detection material of the present invention was used, and the white circles are the results when the standard solution was measured by the conventional method.

As can be seen from this figure, the Cr (VI) concentration and the absorbance show a good proportional relationship (correlation coefficient of 0.999), and Cr (VI) of 5 ppb or more in the sample has 20% of this. By concentrating more than twice, it is possible to measure with high efficiency. Since it is possible to perform visual measurement with a sample of Cr (VI) of 10 ppb or more, if a colorimetric band of the standard solution row of the calibration curve is created in advance, highly toxic Cr ( IV) Cr (VI) can be quantified without using the standard solution.

Example 2 A standard solution for atomic absorption (1000 mg / l) was appropriately diluted with water to prepare a standard solution having a cadmium content of 0 to 0.4 mg / l. 10 ml of each of these cadmium standard solutions was put into a test tube with a lid, and the D obtained in Reference Example 2 was obtained at 5 ° C.
TZ-PNIPAAm was added and shaken to dissolve.
Furthermore, 0.1 ml of 10% NaOH aqueous solution is added, and the temperature is 45 ° C.
Heat for 10 seconds, immediately shake vigorously for 10 seconds,
PNI deposited by incorporating the produced Cd-DTZ complex
PAAm solids were attached to the test tube wall. Then, the liquid portion was removed by decantation, and 1 ml of acetone was added to dissolve the solid portion to obtain a red solution. The result of measuring the absorbance at a wavelength of 535 nm for this solution is shown in FIG.

As can be seen from this figure, the Cd (II) concentration and the absorbance show a good proportional relationship, with 0.0
Cd (II) of 05 mg / l or more can be measured with high efficiency by concentrating it 20 times or more. And 0.
Visual measurement is possible with a sample of 01 mg / l or more. Therefore, if a colorimetric table of a standard solution row is prepared in advance, it is possible to quantify Cd (II) at the time of measurement in the field without using a highly toxic Cd (II) standard solution.

[Brief description of drawings]

FIG. 1 is a perspective view of a kit of the present invention.

FIG. 2 is a graph showing the relationship between the concentration of Cr (VI) and the absorbance at a wavelength of 540 nm.

FIG. 3 is a graph showing the relationship between the concentration of Cd (II) and the absorbance at a wavelength of 535 nm.

[Explanation of symbols]

1 case 2, 4, 5, 6 test tube 3 Pretreatment liquid 7 small bottles 8 dropper 9 Colorimeter 10 Colorimetric band

   ─────────────────────────────────────────────────── ─── Continued front page    F-term (reference) 2G042 BC11 DA06 DA08                 2G054 AB07 CA10 CE01 EA06

Claims (7)

[Claims] 1. A step of: (a) mixing an environmentally-collected sample containing trace contaminants with water, stirring well, allowing to stand, collecting the supernatant, and mixing with a pretreatment liquid; (b) A step of maintaining the above-mentioned mixed liquid at an arbitrary temperature, adding a thermoreversible thermosensitive polymer compound of a fine fibrous solid which becomes hydrophilic at a low temperature having a transition temperature higher than that temperature and becomes hydrophobic at a high temperature, (C) A step of raising the temperature of the solution above the transition temperature of the thermosensitive polymer compound to form a solid, and then separating a liquid portion from the solid; A thermoreversible thermosensitive polymer compound that is hydrophilic at low temperatures and hydrophobic at high temperatures is mixed with a thermoreversible thermosensitive polymer compound in the form of a solid that encloses a hydrophobic coloring reagent that reacts with contaminants and is mixed at a temperature lower than its transition temperature. To form a solution, (e) The temperature of the solution above is raised above the transition temperature,
A step of separating from a liquid portion a solid portion composed of the above thermosensitive polymer compound deposited by inclusion of a coloring compound generated by the reaction of a trace contaminant and the above coloring reagent;
(E) Dissolving the solid part obtained in (b) in water or a solvent at a temperature lower than the transition temperature of the thermosensitive polymer compound to form a solution, and (g) measuring the absorbance of the obtained solution, or A method for quantifying a trace amount of environmental pollutants, which comprises the step of determining the content of a trace pollutant by colorimetrically quantifying a standard product. 2. The method for quantifying environmental pollutant according to claim 1, wherein the trace pollutant in the step (a) is a heavy metal. 3. The method for quantifying trace environmental pollutants according to claim 1, wherein the trace pollutants in the step (a) are harmful inorganic or organic substances. 4. A hydrophobic chelate-forming reagent, which is colored by reacting with a trace contaminant in step (d), is a hydrophobic chelate-forming reagent which forms a chelate compound with a heavy metal ion and is colored. Method for quantifying trace substances of environmental pollution. 5. A thermoreversible type (A) a test tube containing a required amount of pretreatment liquid and having a marked line for adding a sample, and (B) a hydrophilic amount at a required amount of low temperature and hydrophobic at an elevated temperature. Test tube containing thermosensitive polymer, (C) after treatment (B)
A test tube for receiving a liquid portion separated from a hydrophilic-hydrophobic thermoreversible polymer compound solid adhered to the tube wall of
(D) A necessary amount of a thermoreversible polymer compound which is hydrophilic at a low temperature and which is hydrophobic at a high temperature, which contains a hydrophobic coloring reagent which is colored by reacting with a trace contaminant to be quantified, and (C) a test tube with a marked line for adding a liquid portion in (C), (E) a small container containing a solvent for dissolving solid matter adhered to the tube wall of (D) after the above treatment, and (F) A kit for quantifying environmental pollutant trace substances, comprising a colorimeter printed with a colorimetric band corresponding to the concentration of trace pollutants determined by a standard reagent. 6. The thermoreversible thermosensitive polymer compound which is hydrophilic at low temperature and hydrophobic at high temperature in (B) is poly (N-isopropylacrylamide), and (D) is a trace contaminant to be quantified. Is a chromium, and a thermoreversible polymer compound that is hydrophilic at low temperature and is hydrophobic at high temperature in which a hydrophobic color-forming reagent that reacts with it is included in poly (N-isopropylacrylamide) ), And the solvent of (E) is acetone. 7. The thermoreversible thermosensitive polymer compound (B), which is hydrophilic at low temperature and hydrophobic at high temperature, is poly (N).
-Isopropylacrylamide), and (D) a trace amount of environmental pollutant to be quantified is cadmium, which is a thermoreversible type that becomes hydrophilic at low temperature and becomes hydrophobic at high temperature by including a hydrophobic coloring reagent that develops color by reacting with it. The quantitative kit according to claim 5, wherein the polymer compound is poly (N-isopropylacrylamide) including dithizone, and the solvent of (E) is acetone.