WO2004083168A1

WO2004083168A1 - Novel compound suitable for use in chlorine concentration measurement or the like

Info

Publication number: WO2004083168A1
Application number: PCT/JP2004/001634
Authority: WO
Inventors: Ryo Sakamoto; Daikichi Horiguchi
Original assignee: Dojindo Laboratories
Priority date: 2003-02-17
Filing date: 2004-02-16
Publication date: 2004-09-30
Also published as: JP4477578B2; JPWO2004083168A1

Abstract

A compound which has excellent solution stability and is suitable for use as, e.g., a reagent for measurement of the concentration of residual chlorine in water. This compound is represented by the following formula (II) or (III), wherein X is a group represented by the left side in the formula (I), in which the right side is X-. In the formula (I), R1, R2, R3, and R4 each independently is selected among hydrogen, methyl, and methoxy; R5 and R6 each independently is selected among hydrogen and lower alkyls; and R7, R8, R9, R10, and R11 each independently is selected among hydrogen, sulfo, and sulfoalkoxy, provided that the alkyl chain of the sulfoalkoxy may have hydroxy added thereto.

Description

Specification

New compounds suitable for use in chlorine concentration measurement, etc.

The present invention relates to a novel organic compound, and in particular, to a novel compound suitable for use in a measurement system based on an oxidation reaction in an aqueous liquid, such as measurement of a residual chlorine concentration in water. Background art

Water used for drinking and bathing in tap water, swimming pools, baths, etc. uses chlorine agents for the purpose of sterilization. Chlorinic acid exhibits bactericidal action as hypochlorous acid in water. The concentration (free residual chlorine concentration) becomes gradually lower than immediately after addition. Therefore, in order to maintain the bactericidal effect, a free residual chlorine concentration that is always above a certain level is required. For example, it is said that a free residual chlorine concentration of at least 0.2 ppm to 0.4 ppm is required to kill Legionella bacteria in bath water.

Conventionally, o-tolidine, which has been used for measuring residual chlorine concentration in tap water, cannot be used due to its carcinogenicity. As an alternative, DPD (N, N-Jetyl-p-phenylenediamine) is used, but it is commercially available as a powder because it has low stability in solution and must be prepared each time before use. In addition, DPD is not very high, and furthermore, DPD gradually reacts with bound chlorine during the measurement and becomes darker in color, so that the stability of data at the time of measurement is poor.

Under the circumstances described above, in order to measure the residual chlorine concentration in water, the development of a reagent that is highly water-soluble, reacts with free residual chlorine in water, is stable, and develops a strong color is required. For this purpose, a compound in which a monosulfonic acid-substituted alkyl group has been introduced into o-tolidine has been synthesized and has been proposed for use in measuring free residual chlorine. (WO98 / 49-173). However, since this compound also has low stability in a solution state and cannot be used as it is, attempts have been made to stabilize it by adding a surfactant / chelating agent (Japanese Patent Application Laid-Open No. 2002-35004). 16) Solution stabilization has not yet reached the level of practical use. In addition, a compound obtained by introducing a monosulfonic acid-substituted alkyl group into a leuco triamino trifluoromethane compound can also measure residual chlorine.However, this compound also has a problem of solution stability, and has led to practical use. Not.

All of the above compounds are oxidized and develop a color.Therefore, other measurement systems based on an oxidation reaction in an aqueous liquid, for example, when measuring an oxidation reaction involving peroxidase as a substrate for peroxidase. It is also used for However, as described above, they have poor stability in a solution state and are oxidized during storage before being used in an actual measurement system, so that they cannot be used as highly reliable measurement reagents. was there.

An object of the present invention is to provide a new compound having excellent solution stability and suitable as a measuring reagent for a measuring system based on an oxidation reaction in an aqueous liquid such as a measurement of a residual chlorine concentration in water.

Disclosure of the invention

As a result of repeated studies to solve the above problems, the present inventors succeeded in synthesizing a novel compound having a different chemical structure from those conventionally known, and have derived the present invention.

Thus, the present invention provides the following formula (II) or (III) when the atomic group shown on the left side of the following formula (I) is represented by X— as shown on the right side of the following formula (I). And a compound represented by the formula:

In the formula (I), RR ² , R 3 and R ⁴ are each independently selected hydrogen atoms, methyl groups or methoxy groups, and R ⁵ and R ⁶ are each independently selected hydrogen atoms R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and RU are each independently selected a hydrogen atom, a sulfonic acid group or a sulfonic acid alkoxy group, and a sulfonic acid alkoxy group. A hydroxyl group may be added to the alkyl chain.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the results of measuring the solution stability of the compound of the present invention and the comparative compound.

FIG. 2 shows an absorption spectrum in a residual chlorine measurement using the compound of the present invention.

FIG. 3 shows a calibration curve in the measurement of residual chlorine using the compound of the present invention. FIG. 4 shows the absorption spectrum in the measurement of hydrogen peroxide using the compound of the present invention.

FIG. 5 shows a calibration curve in the measurement of hydrogen peroxide using the compound of the present invention. FIG. 6 shows the chemical structural formula of the comparative compound.

Best mode for implementing

As understood from the formulas (I) and (II) or the formulas (I) and (III), the compound of the present invention has a sulfonic acid group added to a benzidine skeleton or a para-roseaniline skeleton. It is a novel compound having a structure in which a benzyl group is introduced.

As described above, in the formula (I), RR ² , R 3 and R 4 are each independently selected from a hydrogen atom, a methyl group or a methoxy group, but the target compound is a compound of the formula (II) In this case, it is preferable that at least one of R ¹ R ² , R ³ and R ⁴ is a methyl group or a methoxy group because a benzidine derivative serving as a starting material can be easily obtained. On the other hand, when the target compound is represented by the formula (III), R ² , R 3 and R ⁴ are generally hydrogen atoms due to the availability of the pararoseurin derivative as a starting material. Is preferred.

In the formula (I), R ⁵ and R ⁶ are a hydrogen atom or a lower alkyl group, each of which is independently selected, as described above. Here, the lower alkyl group is generally an alkyl group having 1 to 4 carbon atoms, and is preferably a methyl group or an ethyl group.

Further, in the formula (I), R7, R8, R9, Rio and Rii are, as described above, each independently selected from a hydrogen atom, a sulfonic acid group or a sulfonic acid alkoxy group; A hydroxyl group may be added to the alkyl chain of the group. Here, the sulfonic acid alkoxy group generally comprises an alkyl chain having 1 to 6 carbon atoms.

Thus, as preferred examples of the compound of the present invention, the following formulas (A) ヽ (B), (C), (D), (E), (F), (G), (H) or (I) ) Potential Forces Of course, the compounds of the present invention are not limited to these. In the chemical structural formulas shown in the present specification and the drawings, carbon atoms and hydrogen atoms may be omitted according to a conventional expression method.

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The compound of the present invention as described above can be synthesized by devising a known reaction. Briefly, an amine (benzidine derivative or pararoseaniline derivative) and an aldehyde or ketone (sulfonic acid group is added) The compound is obtained by preparing a Schiff base by a condensation reaction with a benzyl group-containing aldehyde or ketone, and then reducing the Schiff base (see Examples 1 to 3 described later).

Each of the compounds of the present invention has high water solubility and, when subjected to an oxidation reaction, produces a dye having a specific absorption wavelength. For example, the maximum absorption wavelengths of the dyes of the compounds of formulas (A) to (I) are as shown in Table 1 below.

table 1

A further feature of the compound of the present invention is that it is extremely excellent in solution stability, and does not substantially change with time in an aqueous solution state (see Example 4 described later). It is presumed that the compound of the present invention has such solution stability because the oxidation reaction due to light is suppressed by the introduction of a benzyl group.

Due to the above characteristics, the compound of the present invention can be put to practical use as a reaction reagent in various measurement systems based on an oxidation reaction in an aqueous liquid, in particular, a stable liquid reagent.

That is, according to the present invention, there is provided a reagent for measuring the concentration of residual chlorine in water, comprising the above compound of the present invention as an active ingredient. In another aspect, the present invention relates to a method for measuring the concentration of residual chlorine in water, in which the compound of the present invention is added to a sample to be measured to form a color and the absorption spectrum is measured. However, the present invention also provides a method including a step of detecting the residual chlorine concentration from the absorbance (see Example 5 described later).

Further, the present invention provides a reagent for measuring an oxidation reaction involving peroxidase containing the compound of the present invention as an active ingredient, and a method for measuring an oxidation reaction involving peroxidase, comprising the steps of: The compound of the present invention in the measurement sample The present invention also provides a method comprising the steps of: adding a color to form a color to measure the absorption spectrum; and detecting the hydrogen peroxide concentration and / or the peroxidase activity from the absorbance (see Example 6 described later).

Example

Hereinafter, examples will be given to further illustrate the features of the present invention, but the present invention is not limited by these examples.

Example 1: Synthesis of compound (A)

3 g (l4.1 mmol) of o-tolidine and 8.8 g (28.3 mmol) of benzyl aldehyde 2,4-disulfonic acid sodium salt were dissolved in 150 ml of methanol, and 0.3 ml of concentrated hydrochloric acid was added thereto, followed by refluxing overnight. After cooling, the precipitated crystals were collected by filtration, washed with methanol, and dried in a vacuum desiccator. To a solution of sodium borohydride (500 mg, 13.2 mmol) in water (20 ml), the extracted crystals were added little by little as a solid, and after the addition was completed, the mixture was stirred for 1 hour. After completion of the reaction, the reaction solution was concentrated, and ethyl alcohol was added to precipitate crystals. The crystals were collected by filtration and dried to obtain 9.6 g of compound A and a compound (A) in a yield of 85%.

Compound (A) was identified by NMR, MS and elemental analysis.

1. NMR (300MHz, DMSO-d ₆ )

S 8.1 (d, 2H, J = 2.5Hz), 7.4 (dd, 2H, J = 2.5, 8.0Hz), 7.2 (d, 2H, J = 8.0Hz), 7.l (s, 2H), 7.05 ( d, 2H, J = 9.5Hz), 6.4 (d, 2H, J = 9.5Hz), 5.7 (t, 2H, J = 5.0Hz), 4.7 (d, 4H, J = 6.0Hz), 2.2 (s, 6H)

2. FAB-MS

m / z = 80l [M + H] +

3. Elemental analysis (as octahydrate)

Theory: C, 35.59%; H, 4.27%; N, 2.96%; Na, 9.73%; 0, 33.87%; S, 13.58% Found: C, 35.83%; H, 4.48%; N, 2.76%; Na, 9.62%; 0, 34.10%; S, 13.21% Example 2: Synthesis of compound (C) 2,, 4, 10 g of dihydroxyacetophenone (65.7 thigh) were dissolved in 150 ml of dimethylformamide, and 5.26 g (131.4 mmol) of sodium hydroxide dissolved in 50 ml of water was added little by little. , 3-Propane sultone 17. lg (I40mmol) was added, and the mixture was reacted at 70 ° C for 4 hours. After cooling, the precipitated crystals were collected by filtration and dried to obtain 18 g of 3- [2-acetyl-5- (3-sulfopropoxy) -phenoxy] -1-propane-1-sulfonic acid sodium salt of an intermediate (yield 62). %).

3 g (l4.1 mmol) of o-tolidine and 15.5 g (35.3 mmol) of the intermediate obtained above were dissolved in 200 ml of methanol, and 0.5 ml of concentrated hydrochloric acid was added thereto, and the mixture was refluxed. After cooling, the precipitated crystals were collected by filtration and dried in a desiccator under reduced pressure. Was. After completion of the reaction, the reaction solution was concentrated, and ethyl alcohol was added to precipitate crystals. The crystals were collected by filtration and dried to obtain 11.9 g of compound (C) in a yield of 81%.

Compound (C) was identified by NMR, MS and elemental analysis.

1. NMR (300MHz, DMSO-de)

δ 7.2-7.3 (m, 4H), 7.l (d, 2H, J = 10.0Hz), 6.5 (s, 4H), 6.4 (d, 2H, J = 10.0Hz), 4.3 (s, 2H), 4.15 (t, 4H, J = 8.0Hz), 4.0 (t, 4H, J = 8.0), 2.3-2.5 (m, 8H), 2.2 (s, 6H) _f 1.9-2.l (m, 8H), 1.5 (s, 6H)

2. FAB-MS

m / z = 1062 [M + H] ⁺

3. Elemental analysis (as octahydrate)

Theory: C, 41.86%; H, 5.69%; N, 2.32%; Na, 7.63%; 0, 31.86%; S, 10.64% Found: C, 41.61%; H, 6.02%; N, 2.22%; Na, 7.61%; 0, 31.54%; S, 11.00% Example 3: Synthesis of Compound (H)

Dissolve 5 g (46.3 mmol) of pararoseaniline in 200 ml of methanol and add 11.1 ml of 5 M aqueous sodium hydroxide solution and 1.78 g (47 mmol) of sodium borohydride. At room temperature. After completion of the reaction, the reaction solution was neutralized with hydrochloric acid, and excess sodium borohydride was immersed in the solution.Benzaldehyde monosodium 2,4-disulfonate 46 g (48.2 mmol) and concentrated hydrochloric acid 0.2 nil Was added and refluxed. The precipitated crystals were collected by filtration, dried, dissolved in 50 ml of water, added with 2.8 g (74 mmol) of sodium borohydride, and reacted for 2 hours. After completion of the reaction, the reaction solution was concentrated, and ethyl alcohol was added to precipitate crystals. The crystals were collected by filtration and dried to obtain 40.5 g of a secondary amine compound as an intermediate in a yield of 75%. 10 g (8.5 mmol) of the obtained intermediate was dissolved in 50 ml of methanol. The reaction solution was concentrated, and the concentrate was recrystallized with water-alcohol, and the precipitated crystals were collected by filtration. The obtained crystals were dried under vacuum to obtain 6.1 g of compound (H) in a yield of 57%.

Compound (H) was identified by NMR, MS and elemental analysis.

1. NMR (300MHz, DMSO-d ₆ )

δ 8.l (d, 3H, J = 2.5Hz), 7.4 (dd, 3H, J = 2.5, 8.0Hz), 7.2 (d, 3H, J = 8.0Hz), 7.2 (d, 6H, J = 9.5 Hz), 7.05 (d, 6H, J = 9.5Hz), 5.3 (s, 1H), 4.7 (s, 6H), 3.6 (q, 6H, J = 10.0Hz), 1.7 (t, 9H, J = 10.0 Hz)

2. FAB -MS

m / z = 1257 [M + H] ⁺

3. Elemental analysis (as dodecahydrate)

Theory: C, 37.52%; H, 4.59%; N, 2.85%; Na, 9.37%; 0, 32.60%; S, 13.07% Found: C, 37.58%; H, 4.29%; N, 2.86%; Na, 10.11%; 0, 32.06%; S, 13.28% Example 4: Solution stability of compounds (A) to (I)

Compounds (A), (B), (C), (D), (E), (F), (G), (H), and (I) are dissolved in pure water to 20 mM each. The samples were allowed to stand indoors, and the absorbance at each maximum absorption wavelength shown in Table 1 was measured every day. For comparison, DPD, an o-trisine monosulfonic acid-substituted alkyl group compound (SAT-3: a compound disclosed in Patent Document 2) and a leukotriaminotritrifluoroenyl methane monosulfonic acid-substituted compound are shown for comparison. The same experiment was conducted with an alkyl group compound (TPM-PS). The chemical structural formulas of these comparison compounds are shown in FIG. Fig. 1 shows the changes with time between the seven words. In the comparative compounds, a clear change in absorbance with time was observed, whereas the compounds (A) to (I) of the present invention showed little increase in absorbance for 7 days and were very stable in solution. I understand.

Example 5: Measurement of free residual chlorine using compound (A)

2.0M, 1.0M, 0.5M, 0.25M, 0.125M, 0.0625M Compound (A) solution 45 // 1 was added to develop color. After color development, the absorption spectrum at 400 nm to 800 nm was measured immediately. Fig. 2 shows the spectrum when color was developed with free residual chlorine, and Fig. 3 shows the calibration curve at 675 nm. As a result, it was found that the absorbance at 675 nm increased according to the concentration of hypochlorous acid. The calibration curve showed good linearity.

Example 6: Measurement of hydrogen peroxide using compound (H)

Compound (H) was dissolved in 3 ml of 50 mM phosphate buffer (pH 7.0) to a concentration of lmM and peroxidase to a concentration of 3.3 U / ml. 0 to: An aqueous solution of l.5 mM hydrogen peroxide was applied in an amount of 201, and the color was developed at 20 ° C for 5 minutes, and then the absorption spectrum at 300 to 700 nm was measured. Fig. 4 shows the spectrum when color was developed with peroxidase / hydrogen peroxide, and Fig. 5 shows the calibration curve at 583 nm. As a result, it was found that the absorbance at 583 nm increased according to the concentration of hydrogen peroxide. The calibration curve showed good linearity.

Industrial applicability

As described above, the compound of the present invention produces a dye having a specific absorption wavelength when subjected to oxidizing conditions, but is a very stable new compound when stored in an aqueous solution.

Therefore, the compound of the present invention is oxidized in aqueous liquid based on such properties. As a measurement reagent in a measurement system based on reaction, particularly as a stable and highly reliable liquid reagent, it can be used, for example, for measurement of residual chlorine concentration in water—for measurement of an oxidation reaction involving peroxidase.

Claims

Claims i. The atomic group shown on the left side of the following formula (I) is replaced with X— as shown on the right side of the following formula U), and xx is represented by the following formula (II) or (III). The compound represented.

[In the formula (I), Ri, R ² , R 3 and R ⁴ are each independently selected, a hydrogen atom, a methyl group or a methoxy group, and R ⁵ and R ⁶ are each independently selected. R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are each independently selected hydrogen atom, a sulfonic acid group or a sulfonic acid group, and sulfonic acid A hydroxyl group may be added to the alkyl chain of the alkoxy group. ]

2. The following formula (A), (B), (C) ヽ (D), (E), (F), (G), (H) or (I) The compound according to claim 1, which is represented by any of the following.

3. A reagent for measuring the residual chlorine concentration in water, comprising the compound of claim 1 or 2 as an active ingredient.

4. A reagent for measuring an oxidation reaction involving peroxidase, comprising the compound of claim 1 or 2 as an active ingredient.

5. A method for measuring the residual chlorine concentration in water, wherein the compound according to claim 1 or 2 is added to the sample to be measured to form a color, the absorption spectrum is measured, and the absorbance of the spectrum is measured. Detecting the residual chlorine concentration from the mixture.

6. 'A method for measuring an oxidation reaction involving peroxidase, comprising adding the compound of claim 1 or 2 to the sample to be measured to form a color, measuring the absorption spectrum, and measuring the absorption spectrum. Detecting a hydrogen peroxide concentration and / or a peroxidase activity from the absorbance of the torr.