JP3327241B2 - Reagents and methods for measuring hydrogen sulfide - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a compound of the general formula

Embedded image (Wherein, R ¹ and R ² are a substituted or unsubstituted C ₂ -C ₆ alkyl group, R ³ and R ⁴ are a hydrogen atom, C ₁ -C ₆
It is an alkyl group or a halogen atom. ) And a reagent for measuring hydrogen sulfide, comprising an oxidizing agent,
And a method for measuring hydrogen sulfide using the reagent.

[0002]

2. Description of the Related Art Hydrogen sulfide is a colorless and rotten gas which is naturally contained in volcanic gas, mineral spring water and the like, and is also produced by the decay of sulfur-containing proteins. Industrially, the released hydrogen sulfide contained in exhaust gas, waste liquid, and the like from factories and the like may be oxidized in the atmosphere to form sulfur dioxide gas. Hydrogen sulfide and sulfur dioxide are gases harmful to the human body, and sulfur dioxide causes acid rain and other problems in the environment, so its concentration is regulated. Therefore, it is useful to measure the concentration of hydrogen sulfide in the atmosphere, rivers, soils, factory exhaust gas, factory wastewater, and the like, in order to live a healthy life.

As described above, it is environmentally useful to determine hydrogen sulfide, and its concentration has been measured by various methods. Examples of the measuring method include a method using lead acetate paper or a detector tube, a colorimetric method such as an iodine reduction titration method and a methylene blue method (Talanta, (4) p. 227 (1)
960)).

[0004]

However, in the past, when quantifying hydrogen sulfide in the atmosphere, exhaust gas or waste liquid, a large amount of sample was required because the detection sensitivity was insufficient with the conventional method. The sampling operation was troublesome. Therefore, an object of the present invention is to provide a method for measuring hydrogen sulfide which is simpler and more sensitive than conventional methods for determining hydrogen sulfide.

[0005]

Means for Solving the Problems As a result of intensive studies, the present inventors have found a reagent for measuring hydrogen sulfide comprising the aniline derivative represented by the above general formula (I) and an oxidizing agent. completed. In addition, the present invention provides a method for measuring hydrogen sulfide in a sample obtained by reacting the aniline derivative represented by the general formula (I) with an oxidizing agent in a solution under an acidic condition and improving the colorimetric measurement of the solution by a methylene blue method. Provide the law.

In the measurement of hydrogen sulfide of the present invention, the aniline derivative represented by the general formula (I) is reacted with a solution containing hydrogen sulfide under an acidic condition, and then reacted with an oxidizing agent. It is a method of measuring. Aniline derivative represented by the above general formula (I) are compounds wherein R ¹ and R ² is a substituted or unsubstituted C ₂ -C ₆ alkyl group, C ₂
-C ₆ Preferable examples of the alkyl group include an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, an isopropyl group, isobutyl group. Furthermore, this R
The alkyl group of ¹ and R ² is preferably a C ₃ -C ₄ alkyl group for performing high-sensitivity measurement of hydrogen sulfide and shortening the measurement time. Further, it also is the C ₂ -C ₆ alkyl group having a substituent, examples of the substituent, e.g. sulfo group, sulfonate (sodium sulfonate, potassium sulfonate, etc.) group, mosquitoes Rubamoiru group, Substituted amino groups and the like can be mentioned. The C ₂ -C ₆ alkyl group may be substituted with 1 to 4 of the above substituents. The substituted amino group of the C ₂ -C ₆ alkyl group, a primary amino group or secondary amino group substituted by C ₁ -C ₆ alkyl group, for example methylamino group, ethylamino group, propylamino group Butylamino group, pentylamino group, hexylamino, dimethylamino group, diethylamino group, dipropylamino group, methylethylamino group, ethylpropylamino group and the like.

Further, the substituents R ³ and R ⁴ of the aniline derivative represented by the above general formula (I) are a hydrogen atom, C ₁ -C
_{6 is} an alkyl group or a halogen atom. This C ₁ -C ₆
The alkyl group is a linear or branched C ₁ -C ₆ alkyl group, and examples thereof include a methyl group, an ethyl group, a propyl group, a butyl group, a pentyl group, a hexyl group, an isopropyl group and an isobutyl group. Can be. Further, examples of the halogen atom include a fluorine atom, a chlorine atom, a bromine atom and an iodine atom.

The aniline derivative represented by the general formula (I) includes, for example, p-aminodiethylaniline and p-aminodiethylaniline.
Aminodipropylaniline, p-aminodibutylaniline, p-aminodipentylaniline, p-aminodihexylaniline, p-aminodi (3-sulfopropyl) aniline, 4-amino-N, N-bis [2- (N ′, N '
-Dimethylamino) ethyl] aniline, 4-amino-
N, N-bis [2- (N ′, N′-diethylamino) ethyl] aniline, 4-amino-2,6-dibromo-N,
N-diethylaniline, 4-amino-2,6-dibromo-N, N-dipropylaniline, 4-amino-2,6-
Dichloro-N, N-diethylaniline, 4-amino-
2,6-dichloro-N, N-dipropylaniline and the like can be mentioned. These reagents are easily available, but can also be easily produced by subjecting p-nitroaniline as a raw material to an alkylation reaction, a reduction reaction, or the like.
Further, as the oxidizing agent of the present invention, for example, ferric chloride, potassium ferricyanide, ammonium persulfate, potassium persulfate and the like can be used.

In the measuring method of the present invention, the above-mentioned general formula (I)
The aniline derivative represented by, for example, 0.1 mM to 10
mM, preferably 0.5 mM to 5 mM solution,
Excess amount (usually 10 μM to 5 μM) in the measurement solution containing hydrogen sulfide
00 μM, preferably 20 μM to 200 μM). Further, a mineral acid such as hydrochloric acid, sulfuric acid, nitric acid or the like is added to the measurement solution, and the solution is acidified (pH 2.5 or less, preferably pH 2.5 or less).
2.0 or less). This acid is usually 100 mM to 5 M, preferably 150 mM to
A 600 mM aqueous solution can be added. The oxidizing agent is, for example, 0.2 mM to 8 mM, preferably 0.5 mM
An excess of ８8 mM solution (50 μl to 400 μl, preferably 100 μl to 300 μl) can be added.
The reaction can be carried out at room temperature (about 18 ° C. to about 28 ° C.) to around 50 ° C.

The measurement can be carried out by mixing the aniline derivative represented by the above general formula (I), a measurement solution containing hydrogen sulfide, an acid and an oxidizing agent solution for a time sufficient for the reaction to proceed. Further, an aniline derivative can be added to the measurement solution, and then a mixed solution of an acid and an oxidizing agent can be added. The aniline derivative, the acid and the oxidizing agent represented by the general formula (I) can be constituted as reagents in a kit independently or by mixing two or more kinds.

In carrying out the measurement, first, the maximum absorption wavelength of the dye formed by the reaction of the aniline derivative represented by the above general formula (I) to be used is determined, and colorimetry at that wavelength is a high sensitivity measurement. Preferred to do.

The measurement method of the present invention can usually measure hydrogen sulfide in a solution. For example, to measure hydrogen sulfide in the atmosphere, sodium, iron, cobalt, cadmium, zinc, silver, etc. After a reaction is performed using a metal or its amine derivative to form a sulfide or complex, the sulfide or complex can be measured by the above method (for example, Analyst, Sept., 1974, Vol. 99, p577). -579).

Further, since the measuring method of the present invention can measure a trace amount of hydrogen sulfide produced by enzymatic decomposition of a sulfur-containing amino acid in a living body, it can be used for measuring biological components and the like. it can.

Further, a stabilizer (cyclodextrin, methylated cyclodextrin, etc.) can be added to the reagent of the present invention.

[0015]

The present invention will be described in more detail with reference to the following Examples and Reference Examples.

Reference Example 1 Measurement of hydrogen sulfide by methylene blue method 2 ml of aqueous solution of sodium sulfide (12.5, 25, 50 or 100 μM) in 10 ml of 0.5 g / dl aqueous solution of zinc acetate
l was added. Next, 4 ml of a 75 mg / dl 4N-sulfuric acid solution of p-aminodimethylaniline was added to this solution, and the mixture was left for 5 minutes. Then, ferric chloride 1.2 g / dl 0.0
Add 1 ml of 15M sulfuric acid solution, leave for another 5 minutes,
Hydrogen sulfide was measured from the absorbance at 65 nm. In addition, an aqueous solution to which sodium sulfide was not added was added for measurement. The result is shown in FIG.

Example 1 Measurement of hydrogen sulfide using p-aminodiethylaniline 1 m of an aqueous solution containing 1 mM p-aminodiethylaniline
1N-hydrochloric acid (50 μl) was added thereto, and the sample 2 containing sodium sulfide (12.5, 25, 50 or 100 μM) was added thereto.
00 μl was added. Further, 200 μl of a 10 mM potassium ferricyanide aqueous solution was added, the mixture was kept at 37 ° C. for 10 minutes, and the absorbance at a wavelength of 672 nm was determined to measure hydrogen sulfide. Also,
An aqueous solution to which no sodium sulfide was added was added for measurement. The result is shown in FIG.

Example 2 Measurement of hydrogen sulfide using p-aminodi (3-sulfopropyl) aniline To 1 ml of an aqueous solution containing 1.5 mM p-aminodi (3-sulfopropyl) aniline was added 50 μl of 1N hydrochloric acid.
To this was added 200 μl of a sample containing sodium sulfide (25, 50 or 100 μM). Further, 100 μl of a 10 mM aqueous solution of potassium ferricyanide was added, the mixture was kept at 37 ° C. for 10 minutes, and the absorbance at a wavelength of 675 nm was determined to measure hydrogen sulfide. In addition, an aqueous solution to which sodium sulfide was not added was added for measurement. The result is shown in FIG.

Example 3 Measurement of hydrogen sulfide using p-aminodipropylaniline To 1 ml of an aqueous solution containing 1.5 mM p-aminodipropylaniline was added 50 μl of 1N hydrochloric acid, and sodium sulfide (25, 50 or 100 μM) was added thereto. 200) containing
μl was added. Further, 100 μl of a 10 mM aqueous solution of potassium ferricyanide was added, and the mixture was incubated at 37 ° C. for 10 minutes.
The absorbance was determined at 5 nm, and hydrogen sulfide was measured. In addition, an aqueous solution to which sodium sulfide was not added was added for measurement. FIG. 4 shows the results.

Example 4 Measurement of hydrogen sulfide using p-aminodibutylaniline An aqueous solution containing 1.5 mM p-aminodibutylaniline
50 μl of 1N hydrochloric acid was added to 1 ml, and 200 μl of a sample containing sodium sulfide (25, 50 or 100 μM) was added thereto.
l was added. Further, 100 μl of a 10 mM aqueous solution of potassium ferricyanide was added thereto, and the mixture was kept at 37 ° C. for 10 minutes.
The absorbance was determined in nm and hydrogen sulfide was measured. In addition, an aqueous solution to which sodium sulfide was not added was added for measurement. The result is shown in FIG.

Example 5 4-Amino-N, N-bis [2
Measurement of hydrogen sulfide using-(N ', N'-diethylamino) ethyl] aniline 1.5 mM 4-amino-N, N-bis [2- (N',
[N'-diethylamino) ethyl] aniline (1 ml) is added to 1 ml of an aqueous solution containing 1N hydrochloric acid, and a sample 100 containing sodium sulfide (12.5, 25 or 50 μM) is added thereto.
μl was added. Further, 200 μl of a 10 mM aqueous solution of potassium ferricyanide was added, and the mixture was incubated at 37 ° C. for 10 minutes.
The absorbance at 0 nm was determined, and hydrogen sulfide was measured. In addition, an aqueous solution to which sodium sulfide was not added was added for measurement. FIG. 6 shows the result.

Example 6 Determination of hydrogen sulfide using 4-amino-2,6-dibromo-N, N-dipropylaniline 1.5 mM 4-amino-2,6-dibromo-N, N-
The measurement was carried out in the same manner as in Example 5 using 1 ml of an aqueous solution containing dipropylaniline. In addition, an aqueous solution to which sodium sulfide was not added was added for measurement. FIG. 7 shows the result.

[0023]

According to the present invention, an extremely small amount of hydrogen sulfide contained in a sample can be measured simply and with high sensitivity by using a reagent of the aniline derivative represented by the general formula (I). The sensitivity of the measurement method of the present invention was improved 2 to 18 times as compared with the measurement method of hydrogen sulfide by the methylene blue method.

[Brief description of the drawings]

FIG. 1 is a view showing a result of measuring hydrogen sulfide by a conventional methylene blue method.

FIG. 2 is a view showing a measurement result of hydrogen sulfide using p-aminodiethylaniline.

FIG. 3 is a graph showing the results of measuring hydrogen sulfide using p-aminodi (3-sulfopropyl) aniline.

FIG. 4 is a view showing the measurement results of hydrogen sulfide using p-aminodipropylaniline.

FIG. 5 is a diagram showing the measurement results of hydrogen sulfide using p-aminodibutylaniline.

FIG. 6: 4-Amino-N, N-bis [2- (N ′,
It is a figure which shows the measurement result of hydrogen sulfide using [N'-diethylamino) ethyl] aniline.

FIG. 7 is a graph showing the results of measuring hydrogen sulfide using 4-amino-2,6-dibromo-N, N-dipropylaniline.

Claims (8)

(57) [Claims] 1. A compound of the general formulaWater consisting of an aniline derivative represented by the formula and an oxidizing agent
Elemental measurement reagent (wherein, R ¹ And R^Two Is substituted or unsubstituted
C_Two ~ C₆ An alkyl group;^ThreeAnd R^FourIs hydrogen field
Child, C₁~ C₆An alkyl group or a halogen atom
You. ). 2. The measuring reagent according to claim 1, wherein the substituent on the C ₂ -C ₆ alkyl group of R ¹ and R ² is a sulfo group, a sulfonate group, a carbamoyl group or a substituted amino group. 3. The measuring reagent according to claim 2, wherein the substituent on the C ₂ -C ₆ alkyl group of R ¹ and R ² is a substituted amino group. 4. The measuring reagent according to claim 1, wherein the oxidizing agent is ferric chloride, potassium ferricyanide, ammonium persulfate or potassium persulfate. 5. A compound of the general formula A method for measuring hydrogen sulfide in a sample, comprising reacting an aniline derivative represented by with an oxidizing agent in a solution under acidity and colorimetrically measuring the solution (where R ¹ and R ² are substituted or an unsubstituted C ₂ -C ₆ alkyl group, R ³ and R ⁴ are hydrogen atom, C ₁ -C ₆ alkyl group or a halogen atom.). 6. The method according to claim 5, wherein the substituent on the C ₂ -C ₆ alkyl group of R ¹ and R ² is a sulfo group, a sulfonate group, a carbamoyl group or a substituted amino group. 7. The method according to claim 6, wherein the substituent on the C ₂ -C ₆ alkyl group of R ¹ and R ² is a substituted amino group. 8. The method according to claim 5, wherein the oxidizing agent is ferric chloride, potassium ferricyanide, ammonium persulfate or potassium persulfate.