WO2008012950A1

WO2008012950A1 - Composition for alkylation, and method for detoxification of toxic compound using the composition

Info

Publication number: WO2008012950A1
Application number: PCT/JP2007/000797
Authority: WO
Inventors: Koichiro Nakamura; Akihiro Hishinuma; Shinji Kamiya
Original assignee: Nippon Sheet Glass Company, Limited
Priority date: 2006-07-26
Filing date: 2007-07-26
Publication date: 2008-01-31

Abstract

Disclosed are: a composition useful for the detoxification of a toxic compound containing arsenic or the like efficiently and systematically; and a method for the detoxification of a toxic compound using the composition. Specifically, disclosed are: a composition for alkylation, comprising a cobalt complex; and a detoxification method characterized in that a toxic compound containing at least one element selected from the group consisting of arsenic, antimony and selenium is alkylated in the presence of the composition to thereby detoxify the toxic compound.

Description

Specification

Alkylation composition and method of detoxifying harmful compounds using the composition

Technical field

[0001] The present invention relates to a composition for alkylation and a method for harming harmful compounds using the composition.

Background art

[0002] Heavy metals such as arsenic, antimony and selenium are substances widely used as industrial materials such as semiconductors, but because they are toxic substances to organisms, they are given to organisms by flowing out into the environment. The impact is concerned.

Conventionally, as a method for removing these heavy metals, a coagulant such as polyaluminum chloride (PAG) is added to a wastewater containing inorganic arsenic such as toxic arsenous acid, and the coagulant and iron in raw water are subjected to arsenic. It is generally known a method of aggregation, adsorption, precipitation, removal by filtration, a method of adsorbing an arsenic compound or the like with activated alumina, a cerium-based adsorbent, and the like.

[0004] On the other hand, it has become clear that in the natural world, inorganic arsenic is accumulated in marine organisms such as seaweed, and part of the inorganic arsenic is converted to organic arsenic compounds such as dimethylated arsenic by physiological reaction. (Non-Patent Document 1: Kais et al., 1998, Organomet. Chem. 12 137-143). And, these organic arsenic compounds are generally known to exhibit lower toxicity to mammals than inorganic arsenic.

[0005] Non-Patent Document 1: Kaise et al., 1998, Organomet. Chem. 12 137-143.

Disclosure of the invention

Problem that invention tries to solve

However, in the above method of removing heavy metals using filtration, adsorption, etc., sludge containing harmful compounds such as inorganic arsenic which still remains harmful, and adsorbents on which the harmful compounds are adsorbed are used. , The harmful compound has leaked to the outside In addition, it is necessary to seal or seal with concrete, etc. before storage or burying, and a large storage space and a large space for landfills are required, so there was a problem that mass treatment is difficult.

[0007] In addition, arsenic contained in fish and shellfish is internationally recognized to be non-toxic arsenobetaine, and in the present invention, highly toxic inorganic arsenic is chemically converted to non-toxic arsenobetaine. It is possible to achieve detoxification.

Therefore, in order to solve the above problems, the present invention provides a composition useful for systematically harmonizing harmful compounds containing arsenic etc., and a harmful substance using the composition. The purpose is to provide a method for detoxifying a compound.

Means to solve the problem

[0009] In order to achieve the above object, the present inventors have methylated harmful compounds including arsenic and the like by a chemical reaction using an organic metal complex having a cobalt-carbon bond, particularly preferably dimethylation, more preferably Tried to trimethylate, and as a result of earnestly examining the methylation reaction of the harmful compound, it came to find the present invention.

That is, the composition for alkylation of the present invention is characterized by containing a cobalt complex.

[001 1] In a preferred embodiment of the composition for alkylation according to the present invention, the harmful compound containing at least one element selected from the group consisting of arsenic, antimony and selenium is alkylated by using the cobalt complex. Specializing in

Further, in a preferred embodiment of the composition for alkylation of the present invention,

And a reducing agent for reducing at least one metal selected from the group consisting of arsenic, antimony and selenium.

In a preferred embodiment of the composition for alkylation according to the present invention, the reducing agent is a substance having an SH group.

Furthermore, in a preferred embodiment of the composition for alkylation according to the present invention, the substance having an SH group is glutathione, reduced glutathione (GSH), cysteine, S -Characterized in that it is at least one selected from the group consisting of adenosyl cysteine, sulfolaf anh, homocysteine and thioglycol.

And a methylating agent having an S-Me group.

[001 6] Further, in a preferred embodiment of the composition for alkylation according to the present invention, the methylation addition factor is at least one selected from the group consisting of methionine and S-adenosylmethionine. It is characterized by

[001 7] Further, in a preferred embodiment of the composition for alkylation of the present invention,

And a buffer solution.

Furthermore, in a preferred embodiment of the composition for alkylation according to the present invention, the pH of the buffer solution is in the range of 5 to 10.

[001 9] Further, in a preferred embodiment of the composition for alkylation of the present invention, the pH of the alkylation composition is less than 9.

, H ₂ 0 ₂ .

And an organic halogen compound.

In a preferred embodiment of the composition for alkylation according to the present invention, the organic halogen compound is a methyl halide.

In a preferred embodiment of the composition for alkylation according to the present invention, the methyl halide is at least one selected from the group consisting of methyl iodide, methyl bromide and methyl chloride. It features.

In a preferred embodiment of the composition for alkylation according to the present invention, the organic halogen compound is a halogenated acetic acid.

Furthermore, in a preferred embodiment of the composition for alkylation according to the present invention, the halogenated acetic acid is at least one selected from the group consisting of crocodile acetic acid, bromoacetic acid, and joud acetic acid. I assume.

In a preferred embodiment of the composition for alkylation according to the present invention, Halogen compounds such as methyl chloride, methyl bromide, methyl iodide, cloroacetic acid, bromoacetic acid, bromoacetic acid, chloroacetic acid, crocodile ethanol, bromoethanol, bromoethanol, chloroethanol, croropropionic acid, bromopropionic acid, It is characterized in that it is at least one selected from the group consisting of monopropionic acid, crochiral acetic acid ethyl ester, bromoacetic acid ethyl ester, and sodium acetic acid ethyl ester.

Further, in a preferred embodiment of the composition for alkylation according to the present invention, the organic halogen compound is represented by the chemical formula 1:

Chemical formula 1 RMgX

It is characterized in that it is a Grignard reagent represented by (wherein R = Me, GH ₂ G00H, or GH ₂ G00 G ₂ H ₅ in Chemical Formula 1; X = G and Br or I).

Furthermore, in a preferred embodiment of the composition for alkylation according to the present invention, the organic halogen compound is selected from the group consisting of: pesticides, flame retardants, dioxins, PGB, DDT, trihalomethanes, trichloroethyls, croforms. It is characterized by being derived from persistent organic substances.

In a preferred embodiment of the composition for alkylation of the present invention, the composition further comprises a reducing agent for reducing a cobalt complex.

In a preferred embodiment of the composition for alkylation according to the present invention, the reducing agent is at least one selected from the group consisting of titanium oxide or ruthenium complex.

Further, in a preferred embodiment of the composition for alkylation according to the present invention, the cobalt complex is methylcobalamin (methylated vitamin B 12, formal name: Go —-[5, 6-dimethylbenz] H 1-Imidazoyl 1-yl 1 Go S-methylcobamide], vitamin B 12 such as cyanocobalamin, cobalt (II) acetate, cobalt (III) acetylacetonate Cobalt force Ruponyl (Nicobalt octacarponyl), Cobalt (11) 1, 1, 1, 5, 5, 5-Hexafluoroacetylacetonato, Cobalt (II) Meso-1 tetrapheniol porphyrin, Hexafluorophosphate bis (pentamethylcyclopentagenyl ) Cobalt, N, Ν '— bis (salicylidene) ethylene diamine cobalt (I

I), bis (2,2,6,6-tetramethyl-3-, 5_heptanedionate) cobalt (II), (clorophthalocyaninato) cobalt (||), chloro tris (triphenyl phosphine) cobalt (I), Methyl complex of cobalt (II) acetate, cobalt (II) benzoate, cobalt (II) cyanide, cobalt (III)

), 2- (Ethyl) hexanoic acid cobalt (II), meso-tetramethyoxyphenylophylline cobalt (II), cobalt naphthenate, phthalocyanine cobalt (

II), methylcobalt (III) protoporphyrin IX, cobalt stearate, cobalt (II) sulfamate, (1R, 2R) _ (_) _1, 2-cyclohexanediamino-, Ν '-bis (3) , 5-di_t_ putyl salicylidene) cobalt (II), (1 S, 2S) _ (+) _1, 2-cyclohexanediamino-, Ν '-bis (3, 5-di _ t_ petitulicylidene) ) Cobalt (II), cyclopentajenyl bis (triphenyl phosphine) cobalt (I), cyclopentajenyl cobalt dicarpoyl, dibromobis (triphenyl phosphine) cobalt (ll), (tetraaminochlorophthalocyaninato) cobalt (II), (Tetramethyl t-butyl phthalocyaninato) Methyl complex of at least one compound selected from cobalt (II), or the cobalt compound and halogen It is characterized in that it is at least one selected from the group consisting of alkyl halides, in particular cobalt monomethyl complexes formed by coexisting methyl halides.

Further, in a preferred embodiment of the composition for alkylation according to the present invention, a molar concentration of the reducing agent for reducing at least one metal selected from the group consisting of arsenic, antimony and selenium [Reducing Agent] It is characterized in that it is a ratio with a metal concentration [Metal] of a metal selected from Arsenic, Antimony and Selenium, that is, [Reducing Age] / [Metal] force is 1 000 or more.

In a preferred embodiment of the composition for alkylation according to the present invention, the ratio is 10000 or more.

Furthermore, in a preferred embodiment of the composition for alkylation according to the present invention, the molar concentration of the cobalt complex [Go complex], and selected from arsenic, antimony, selenium It is characterized in that the ratio to the molar concentration of metal [Metal], that is, [Go comp l ex] / [Metal] force is 100 or more.

Further, in a preferred embodiment of the composition for alkylation according to the present invention, the ratio is preferably 1000 or more.

In the preferred embodiment of the composition for alkylation according to the present invention, the method for detoxifying harmful compounds according to the present invention comprises the presence of the composition according to any one of claims 1 to 6, It is characterized in that harmful compounds containing at least one element selected from the group consisting of arsenic, antimony and selenium are rendered harmless by alkylation.

[0037] In a preferred embodiment of the harmful compound detoxification method of the present invention, the method is characterized in that the valence number of one of the elements is a high oxidation number to render it harmless.

[0038] Further, in a preferred embodiment of the method for detoxifying harmful compounds according to the present invention, at least one bond of one of the elements described above is alkylated.

[0039] In a preferred embodiment of the method for making harmful compounds harmless according to the present invention, the above-mentioned element is arsenic.

Further, in a preferred embodiment of the harmful compound detoxification method of the present invention, the 50% lethal dose (LD ₅₀ ) of the compound detoxified by the alkylation is 10

It is characterized by being at least 00 mg / kg.

Further, in a preferred embodiment of the harmful compound detoxification method of the present invention, the 50% cell growth inhibitory concentration of the compound detoxified by the alkylation as described above (I

C ₅₀ ) is characterized by being 1000 1000 or more.

Further, in a preferred embodiment of the harmful compound detoxification method according to the present invention, the harmful compound is selected from arsenous acid, arsenic pentoxide, arsenic trichloride, arsenic pentachloride, arsenic sulfide compound, cyano-arsenic compound, It is characterized in that it is selected from the group consisting of a nitrogen-containing arsenic compound and other arsenic inorganic salts.

[0043] In addition, in a preferred embodiment of the method for detoxifying harmful compounds of the present invention, Characterized in that the alkylation is methylation.

Further, in a preferred embodiment of the method for making harm to harmful compounds according to the present invention, the harmful compounds are converted into dimethyl compounds or trimethyl compounds by the above-mentioned methylation.

Further, in a preferred embodiment of the harmful compound detoxification method of the present invention, the dimethyl compound is dimethylarsonylethanol (DMAE), dimethylarsonyl acetate (DMAA), dimethylarsinic acid, or arsenosugar It is characterized by being.

[0046] In a preferred embodiment of the harmful compound detoxification method of the present invention, the above-mentioned trimethyl compound is characterized by being an arsenocholine, an arsenobetaine, a trimethylasenosugar, or a trimethyl arsenoxide.

[0047] Further, the method for detoxifying harmful halides of the present invention is a pesticide, a flame retardant, a dioxin, a PGB, a DDT, in the presence of the composition according to any one of the items 1 to 26. The invention is characterized in that the organic halide selected from the group consisting of trihalomethane, trichloroethyl and chloroform is detoxified by dehalogenation.

[0048] In a preferred embodiment of the method of making harmful compounds harmless according to the present invention, a pesticide, a flame retardant, dioxin, in the presence of the composition according to any one of claims 1 to 26. An organic halide selected from the group consisting of PGB, DDT, trihalomethane, trichloroethyl and chloroform is detoxified by dehalogenation, and in the presence of a cobalt complex obtained by the reaction, arsenic, antimony A harmful compound containing at least one element selected from the group consisting of selenium is characterized by rendering it harmless by alkylation.

In a preferred embodiment of the harmful compound detoxification method of the present invention, the method is further characterized by irradiation with light in the presence of a reducing agent that reduces a cobalt complex.

Further, in a preferred embodiment of the method for making harm to harmful compounds according to the present invention, the reducing agent is selected from the group consisting of titanium oxide or ruthenium complex. It is characterized by at least one type.

Effect of the invention

The composition for alkylation of the present invention has an advantageous effect that it is possible to easily and conveniently alkylate harmful compounds, particularly harmful compounds containing arsenic, antimony, selenium and the like. Further, according to the method of the present invention, since harmful compounds can be harmlessly reduced as much as possible, the advantageous effect is obtained that a large space such as a storage place is not required. Further, according to the method of the present invention, since the living object itself is not used alive, there is an advantageous effect that unnecessary by-products are not generated. Furthermore, according to the present invention, it is possible to achieve an advantageous effect that harmful inorganic arsenic and the like can be further reduced by a simple operation.

Brief description of the drawings

[FIG. 1] FIG. 1 is a diagram showing H P L C — I C P-MS analysis (upper: standard sample, lower: sample) of Chlorella extract.

[Fig. 2] Fig. 2 shows HPL C-ICP-MS analysis of the extract of Quailella (upper: standard sample, middle: GSH addition (NEJ4-7), lower: Me Co + GSH + MIAA addition (NEJ5 -7) is a figure which shows.

[FIG. 3] FIG. 3 is a view showing a state in which GSH addition (NEJ4-4) and GSH + MeCo + MIAA addition (NEJ5-4) are added to the Chlorella extract and NOH-treated (bottom).

[FIG. 4] FIG. 4 is a diagram showing a state in which GSH + MeCo + MIAA is added (NE_9_4) to DMA.

[FIG. 5] FIG. 5 is a diagram showing an HPLG-IGP-MS chromatogram. The numbers in the figure correspond to the numbers in Table 7.

[FIG. 6] FIG. 6 shows an HPLG-IGP-MS chromatogram. The numbers in the figure correspond to the numbers in Table 7.

[FIG. 7] FIG. 7 is a view showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. It is a graph of the results of Table 7.

[FIG. 8] FIG. 8 is a view showing the time-dependent change of the proportion of the arsenic compound in the reaction solution. [FIG. 9] FIG. 9 is a view showing the time-dependent change of the proportion of the arsenic compound in the reaction solution.

[FIG. 10] FIG. 10 shows an HPLG-IGP-MS chromatogram. No. in the figure corresponds to the numbers in Table 8.

[FIG. 11] FIG. 11 is a diagram showing an HPLG-IGP-MS chromatogram. No. in the figure corresponds to the numbers in Table 8.

[FIG. 12] FIG. 12 is a diagram showing an HPLG-IGP-MS chromatogram. No. in the figure corresponds to the numbers in Table 8.

[FIG. 13] FIG. 13 is a diagram showing an HPLG-IGP-MS chromatogram. No. in the figure corresponds to the numbers in Table 8.

[FIG. 14] FIG. 14 is a diagram showing the time-dependent change of the concentration of the arsenic compound in the reaction solution.

[FIG. 15] FIG. 15 is a view showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. (After hydrogen peroxide treatment)

[FIG. 16] FIG. 16 is a diagram showing the time-dependent change of the proportion of the arsenic compound in the reaction solution. (Before hydrogen peroxide solution treatment)

[FIG. 17] FIG. 17 is a view showing the time-dependent change of the proportion of the arsenic compound in the reaction solution. (After hydrogen peroxide treatment)

[FIG. 18] FIG. 18 is a view showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. (No. 1 to No. 4 in Table 9)

[FIG. 19] FIG. 19 is a view showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. (No. 5 to 5 in Table 9, after hydrogen peroxide treatment)

[FIG. 20] FIG. 20 is a diagram showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. (No. 9 to 12. 12 in Table 9)

[FIG. 21] FIG. 21 is a diagram showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. (No. 13 to No. 16 in Table 9, after hydrogen peroxide treatment)

FIG. 22 is a diagram showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. (No.17 to No.20 in Table 9, before hydrogen peroxide treatment)

[FIG. 23] FIG. 23 is a diagram showing the time-dependent change of the concentration of the arsenic compound in the reaction solution. . (No. 21 to No. 24 in Table 9, before hydrogen peroxide treatment)

[FIG. 24] FIG. 24 shows a time-dependent change of the proportion of arsenic compounds in the reaction solution. (No. 1 to 4 in Table 9, before hydrogen peroxide treatment)

[FIG. 25] FIG. 25 shows the change with time of the proportion of arsenic compounds in the reaction solution. (No. 5 to 8 in Table 9, after hydrogen peroxide treatment)

[FIG. 26] FIG. 26 shows the change with time of the proportion of arsenic compounds in the reaction solution. (No. 9 to No. 12, Table 9, before hydrogen peroxide treatment)

[FIG. 27] FIG. 27 shows the change with time of the proportion of arsenic compounds in the reaction solution. (No. 13 to No. 16 in Table 9, after hydrogen peroxide treatment)

[FIG. 28] FIG. 28 is a diagram showing the change with time of the proportion of arsenic compounds in the reaction solution. (No. 17 to No. 20 in Table 9, before hydrogen peroxide treatment)

[FIG. 29] FIG. 29 is a diagram showing the time-dependent change of the proportion of the arsenic compound in the reaction solution. (No. 21 to No. 24 in Table 9, after hydrogen peroxide treatment)

[FIG. 30] FIG. 30 is a view showing the mechanism of arsenite methylation in the case of vitamin B12 as an example.

[FIG. 31] FIG. 31 is a diagram showing an H P L C-I C P-M S chromatogram of a methylation reaction of selenite (Se (I V)) by M C.

[FIG. 32] FIG. 32 is a diagram showing an HPLG-I GP-MS chromatogram (elements to be measured: Sb, m / z 121).

[FIG. 33] FIG. 33 is a diagram showing reaction conditions for formation of trimethylarsenic (TMA) from arsenous acid by methylcobalamin.

BEST MODE FOR CARRYING OUT THE INVENTION

The composition for alkylation of the present invention contains a cobalt complex. Here, the cobalt complex is not particularly limited, and examples thereof include organic metal complexes having a cobalt-carbon bond.

The following can be exemplified as an example of the organic metal complex having a cobalt-carbon bond. That is, methylcobalamin (or methylated vitamin B12, formal name: Gohi [hi_5, 6-dimethylbenz 1 1 H-imidazo 1 1-1 1 1 GOyS-methyl cobalt is preferably used. In addition, vitamin B12 such as cyanocobalamin, cobalt (II) acetylacetonate, cobalt (III) acetylacetonato, cobalt carponyl (nicobalt octacalponyl), cobalt (11) 1, 1, 1, 5,5,5-hexafluoroacetylacetonato, cobalt (II) meso-one tetraphenyl porphine, hexafluoromouth phosphate bis (pentamethylcyclopentajenyl) cobalt, N, N '-bis (Salicylidene) ethylenediaminecobalt (II), bis (2, 2, 6, 6-tetramethyl-3, 5-heptanedionato) cobalt (ll), (clorophthalocyaninato) cobalt (||) Chloro tris (triphenyl phosphine) cobalt (I), methyl complex of cobalt (II) acetate, cobalt (II) benzoate, cobalt (II) cyanide, Hexane cobaltate cobalt (II), 2-ethyl cobaltate cobalt (II), meso-tetramethoxyphenyl porphyrin cobalt (II) cobalt naphthenate, phthalocyanine cobalt (I 1), methyl cobalt (III) pro Toporphyrin IX, cobalt stearate, cobalt (II) sulfamate, (1R, 2R) _ (_) _1, 2-cyclohexanediamino-, Ν'- bis (3, 5-di _ t _ ptyl salicylidene) Cobalt (II), (1S, 2S) _ (+) _1, 2-Cyclohexane diamine-Ν, Ν '-Bis (3, 5-di _ t _ pyl salicylidene) cobalt (II), cyclopenta geni (Bis (triphenylphosphine) cobalt (I), cyclopentadienylcobalt dicarponyl, dibromobis (triphenylphosphine) core / relet (||), (tetraaminochlorophthalocyanine) Aninato) methyl complex of at least one compound selected from anthraito (II), (tetra-t-butyl phthalocyaninato) cobalt (II), or the cobalt compound and an alkyl halide, particularly halogenated An example is a cobalt monomethyl complex which is formed in the coexistence of methyl. Methylcobalamin is preferable as the organometallic complex having a cobalt-carbon bond, from the viewpoint that harmful compounds containing harmful inorganic arsenic and the like can be relatively easily alkylated to make the organic matter less toxic.

That is, in the composition for alkylation of the present invention, By using it, it is possible to alkylate harmful compounds containing at least one element selected from the group consisting of arsenic, antimony and selenium. Here, in the present specification, the term "harmful compound" means a compound that may leak to the environment and cause any adverse effect on organisms when exposed to a living thing.

Among the above-mentioned harmful compounds, the harmful compounds containing arsenic include arsenous acid, arsenic pentoxide, arsenic trichloride, arsenic pentachloride, arsenic sulfide compounds, cyano-arsenic compounds, chloroarsenic compounds, and the like. Arsenic inorganic salts etc. are mentioned. These _arsenics , for example, have LD _5Q (mg / kg) (50% lethal dose in mice) of 20 or less, and are generally toxic to organisms.

Further, examples of the harmful compound containing antimony include antimony trioxide, antimony pentoxide, antimony trichloride, antimony pentachloride and the like.

Furthermore, examples of the harmful compound containing selenium include selenium dioxide, selenium trioxide and the like.

[0059] In a preferred embodiment, the composition of the present invention may further contain a reducing agent that reduces at least one metal selected from the group consisting of arsenic, antimony and selenium. Alkylation can be further promoted by the presence of such a reducing agent. The ability to reduce arsenic in conversion of arsenic to arsenobetaine or the possibility of rate-limiting methyl transfer reaction may be considered, but addition of a reducing agent promotes conversion to arsenobetaine etc. It can be considered that As such a reducing agent, for example, a substance having an SH group can be mentioned. Specifically, a substance having an SH group is, for example, glutathione, reduced glutathione (GSH), cysteine, S-adenosylcysteine, Mention may be made of at least one selected from the group consisting of sulphofurafan, homocysteine and thioglycol. Also, any combination of these may be used. For example, glutatin + homocysteine, glutathion + thioglycol can be mentioned.

Further, in a preferred embodiment of the composition, arsenic, antimony, selenium or the like The molar ratio of the reducing agent that reduces at least one metal selected from the group consisting of [Reducing Agent] to the molar concentration of the metal selected from Arsenic, Antimony, Selenium [Metal], ie [Reducing Agent] Agent] / [Metal] is more than 1000. More preferably, the ratio is 10000 or more. When the composition of the present invention is applied to the method for detoxifying harmful compounds, etc. described later, under such conditions, the alkylation is achieved at a high ratio, so that harmful compounds containing arsenic etc. From the perspective of being able to achieve the detoxification of

In a preferred embodiment of the composition for alkylation according to the present invention, the molar concentration [Go complex] of the cobalt complex and the molar concentration [Metal] of a metal selected from arsenic, antimony and selenium The ratio, ie [Go complex] / [Metal] force is over 100. More preferably, the ratio is 1000 or more. When the composition of the present invention is applied to a method for making harmless a harmful compound, etc. described later, alkylation is achieved at a high rate under such conditions, and the harmful compound containing arsenic etc. is obtained. It is from the point of view that detoxification can be achieved efficiently.

According to the molar concentration ratio described above, one of the main objects of the present invention, namely, the toxicity of inorganic arsenic (acute toxicity value: LD50 0.03 g / kg) etc. The purpose is to achieve more efficient conversion to methylated arsenic etc. The target products, low-toxic methylated arsenic and the like, are trimethylalsuoxide (acute toxicity value: LD 50 10.6 g / kg) and arsenobetaine (acute toxicity value: LD50 10. Og / kg). It is possible to reduce the toxicity of 1/300 to inorganic arsenic etc., preferably 10% or more, more preferably 50% or more, still more preferably 90% or more of these nontoxic arsenic etc. It is possible to obtain in relative yields.

Also, in a preferred embodiment of the composition for alkylation of the present invention, the composition further comprises a methylation addition factor having an S-Me group. The presence of the methylating additive can provide more alkyl groups and thus, more Various alkylation and detoxification can be achieved. Examples of the methylation additive include at least one member selected from the group consisting of methionine and S-adenosylmethionine.

In the composition for alkylation of the present invention, a buffer may be contained. As the buffer, one usually used for isolation, purification, storage, etc. of biological materials can be used, and it is not particularly limited. For example, Tris buffer, phosphate buffer, carbonate buffer, A buffer such as borate buffer can be exemplified. The pH value of the buffer solution is preferably in the range of 5 to 10 in consideration of safer detoxification. Further, the pH of the composition for alkylation is more preferably less than 9. The composition for alkylation of the present invention can further contain H ₂ O ₂ . That is, hydrogen peroxide may be added to lower the acute toxicity by increasing the oxidation state (trivalent → pentavalent).

The composition for alkylation of the present invention can further contain an organic halogen compound. From the viewpoint of facilitating the conversion of a dimethyl compound and / or a trimethyl compound to an arsenobenzene, an example of the organic halogen compound is methyl halide. As the methyl halide, at least one selected from the group consisting of methyl iodide, methyl bromide and methyl chloride can be mentioned from the viewpoint of high methylation reactivity.

In addition, the organic halogen compound is selected from the group consisting of sodium acetate, sodium ethanol, bromoacetic acid, bromoethanol and sodium propionic acid from the viewpoint of high alkylation reactivity. At least one can be mentioned.

In a preferred embodiment, the organic halogen compound is a halogenated acetic acid. As an example of the halogenated acetic acid, at least one selected from the group consisting of crocodile acetic acid, bromoacetic acid, and joud acetic acid can be mentioned.

In a preferred embodiment, the organic halogen compounds include methyl chloride, methyl bromide, methyl iodide, crocodile acetic acid, bromoacetic acid, bromoacetic acid, chloroacetic acid, crocodile ethanol, bromoethanol, choloethanol One-handed one, black mouth pro Examples thereof include at least one selected from the group consisting of pyonic acid, bromopropionic acid, iodopropionic acid, acetic acid ester of acetic acid, bromoacetic acid ethyl ester, bromoacetic acid ethyl ester and iodoacetic acid ethyl ester.

Furthermore, in the present invention, the organic halogen compound is represented by the following Chemical Formula 1: Chemical Formula 1 RMgX

It may be a Grignard reagent represented by (wherein R = Me, GH ₂ G00H, or GH ₂ G00G ₂ H ₅ in Chemical Formula 1; X = G and Br or I).

Also, the organic halogen compounds described above are mainly for facilitating the conversion of dimethyl compounds and / or trimethyl compounds into stable arsenobetaines when methylating harmful compounds. I explained the points that can be used from the viewpoint.

The organic halogen compounds exemplified below are those listed in the method for detoxifying the organic halide of the present invention described later which can be the target of detoxification by dehalogenation.

That is, when the organic halogen compounds to be detoxified are listed, it is possible to cite those selected from the group consisting of pesticides, flame retardants, dioxins, PGB, DDT, trihalomethanes, trichloroethyls, chlorforms. it can. When these substances are not purified, they may be introduced into the reaction system by any conventional method such as extraction and separation (whether liquid, gas, solid, etc.). Since a cobalt complex is present in the composition for alkylation of the present invention, the harmful organic halide can be dehalogenated by the catalytic action of the cobalt complex, and thus, dehalogenation leads to an organic compound. The halide can be detoxified.

The composition for alkylation of the present invention may further contain a reducing agent that reduces the cobalt complex. This has the advantageous effect that the presence of the reducing agent makes it possible to change the active cobalt complex to the oxidation state as described later.

Such a reducing agent is not particularly limited as long as the cobalt complex can be activated. For example, at least one selected from the group consisting of titanium oxide or a ruthenium complex can be mentioned, although it is not.

Next, the method for detoxifying harmful compounds of the present invention will be described. That is, in the method for detoxifying harmful compounds of the present invention, a harmful compound containing at least one element selected from the group consisting of arsenic, antimony and selenium in the presence of the above-mentioned composition for alkylation of the present invention is used. Detoxify by alkylating. Here, the composition for alkylation and the harmful compound of the present invention mean those described above and can be applied as they are in the detoxifying method of the present invention.

In a preferred embodiment of the detoxification method of the present invention, 50% cell growth inhibitory concentration

(IC ₅₀ ) or LD ₅₀ is large, and from the viewpoint of achieving further harmlessization, to make the harmful compound harmless by setting the valence number of the one element contained in the harmful compound to be a high oxidation number. Is preferred. Specifically, it is possible to make the valence number of the above one element a high oxidation number by alkylation using the composition of the present invention described above as a catalyst for the reaction. In the case where the above element is arsenic or anthive, it is preferable to set the trivalent valence to pentavalent, and in the case of selenium, the tetravalent valence to hexavalent.

In the present invention, the harmless treatment of the harmful compound is carried out by alkylating the harmful compound. Here, detoxification can be achieved by alkylating at least one bond of the one element in the harmful compound.

Specifically, at least one bond of the above-described one type of element can be alkylated by performing a reaction using the above-mentioned alkylation composition of the present invention. Here, examples of the alkyl group added to the above-mentioned one type of element include a methyl group, a ethyl group, a propyl group and the like. From the viewpoint of achieving detoxification more efficiently, a methyl group is preferred as the alkyl group.

[0079] In the detoxification method of the present invention, 50% lethal dose (LD _M ) (50% of mice is killed) of the compound detoxified by the above-mentioned alkylation from the viewpoint of safety to the living body Oral toxicity due to drug dose) preferred to be 10001000 mg / kg And more preferably 5000 mg / kg or more.

Further, in the detoxifying method of the present invention, from the viewpoint of safety to the living body, the 50% cell growth inhibitory concentration (IG ₅₀ ) of the compound detoxified by the above alkylation or arylation is , 1000; u M or more is preferable, 3000; u M or more is more preferable. Here, as used herein, 50% cell growth inhibitory concentration (IG ₅₀ ) means a numerical value indicating the concentration of a substance necessary to inhibit or inhibit the growth of 100 cells by 50% together with a substance. means. The smaller the IG _5Q value, the greater the cytotoxicity. Here, the IG _M was calculated from the result of examining the cytotoxicity of plasmid DNA damage under conditions of 37 ° C. and 24 hours.

Here, the IG _M of each arsenic compound is shown in Table 1.

[Table 1]

I C 5 0 value (ju M)

2 4 h, 3 7 ^a C

From Table 1, it is found that the arsenosugars having trivalent arsenic are more cytotoxic than monomethylated arsenic (MMA) and dimethylated arsenic (DMA) that have pentavalent arsenic, but trivalent arsenic It is found to be less cytotoxic than MMA, DMA and arsenous acid. On the other hand, MMA with trivalent arsenic and DMA are more cytotoxic than arsenous acid (trivalent and pentavalent), but overall, from the viewpoint of cytotoxicity, arsenic compounds with pentavalent arsenic are more effective. It can be understood that the safety to the living body is higher than the arsenic compound having trivalent arsenic.

Further, LD _5Q of each arsenic compound is shown in Table 2.

[0085] 2) Arsenic chemical species LD (m

A (III) Inorganic Arsenic (Trivalent) 4.5

A (V) Inorganic form Arsenic (pentavalent) 1 4 1 8

MMA monomethylarsonic acid 1, 8 0 0

D MA dimethylarsinic acid 1, 2 0 0

A C Anoresenocholine 6, 0 0 0

T MA O Trimethylarsoxide 1 0, 6 0 0

A B A ^ ^ Cenobetaine 1 0, 0 0 0

Further, in the harmlessization method of the present invention, it is preferable that the biological half life of the compound detoxified by the above alkylation is 8 hours or less from the viewpoint of safety to the living body . In the detoxifying method of the present invention, it is preferable that the harmful compound be a dimethyl compound or a trimethyl compound by the methylation from the viewpoint of safety and low toxicity. Examples of the dimethyl compound include dimethylarsonylethanol (DMAE), dimethylarsonylacetate (DMAA), dimethylarsinic acid, and arsenosugar. In addition, as the trimethyl compound, it is possible to cite arsenocholine, arsenobetaine, trimethylarsenosugar or trimethylarsinoxide.

Further, the method for detoxifying the organic halide of the present invention is selected from the group consisting of pesticides, flame retardants, dioxins, PGBs, DDTs, trihalomethanes, trichloromethanes, chloroform in the presence of the composition of the present invention. It is characterized by detoxifying organic halides by de / arogenation.

Furthermore, in a preferred embodiment of the harmful compound detoxification method of the present invention, in the presence of the above-mentioned composition of the present invention, pesticides, flame retardants, dioxins, PGBs, DDs, trihalomethanes, trichloroethyls, An organic halide selected from the group consisting of cloroform is detoxified by dehalogenation, and the group consisting of arsenic, antimony and selenium in the presence of a cobalt complex obtained by the reaction. It is possible to render harmless by alkylating harmful compounds containing at least one element selected from

That is, in the presence of the composition for alkylation of the present invention, organic halides which are inherently harmful, such as pesticides, flame retardants, dioxins, PGBs, DDTs, trihalomethanes, trichloroethyls, croforms, etc. In the reaction, dehalogenation occurs and the reaction produces an organic cobalt complex, and the organic substance in the organic cobalt complex can be one of the sources of the alkyl group for the alkylation of harmful heavy metals. That is, it is possible to convert harmful compounds, such as inorganic arsenic, into harmless substances, ie, organic arsenic, by using the thus-obtained source of alkyl group.

Furthermore, in a preferred embodiment of the harmful compound of the present invention, light is irradiated in the presence of a reducing agent which reduces a cobalt complex. By irradiation with light, it is possible to change the oxidation state to cobalt complex (I I) force active cobalt complex (I). The cobalt complex (I) is detoxified by dehalogenation by reacting the cobalt complex (I) with the harmful organic halogen compound for dehalogenating the complex, and the cobalt complex is supplied with an alkyl group. It produces the advantageous effect of being able to obtain organic matter that can be a source.

Such a reducing agent is not particularly limited as long as the cobalt complex can be activated, and, for example, at least one selected from the group consisting of titanium oxide or ruthenium complex may be mentioned. it can.

Next, the method for making organic halides harmless according to the present invention is as follows.

That is, the method for detoxifying the organic halide of the present invention comprises, in the presence of the composition of the present invention described above, an agricultural chemical, a flame retardant, dioxin, PGB, DDT, trihalomethane, trichloroethl, croform. It is characterized in that the organic halide selected from the group is detoxified by dehalogenation. Organic halides selected from the group consisting of pesticides, flame retardants, dioxins, PGBs, DDTs, trihalomethanes, trichloroethyls, chloroforms are introduced into the reaction system If it is in a form that can not be introduced, it can be introduced as an appropriate form (whether gas, liquid, or solid) by extraction, separation, purification, etc. by a conventional method. According to this method, the cobalt complex in the composition of the present invention contributes not only to the alkylation but also to the dehalogenation of the organic halide, and also to the detoxification of the organic halide. Can be achieved. Thus, the composition of the present invention is very useful. Further, in the same manner as the harmless method of the harmful compound of the present invention described above, light may be irradiated in the presence of a reducing agent for reducing a cobalt complex. For the explanation of the reducing agent, etc., the explanation of the above-mentioned method of detoxifying harmful compounds can be applied as it is.

Taking vitamin B12 (methylcobalamin: GH ₃ _Gob (III)) as an example, the mechanism for arsenite methylation is as shown in FIG. In Fig. 30, iAs (V) is pentavalent inorganic arsenic, iAs (lll) is trivalent inorganic arsenic, ATG is triglutathione arsenic complex, MADG is monomethyldiglutathione arsenic complex, DMAG is dimethyl glutathione arsenic complex It is an abbreviation of

Example

Examples of the present invention will be described below, but the following examples do not in any way limit the scope of the present invention.

First, abbreviations used in the examples are as follows.

[Abbreviation]

i A s (I I I): Inorganic trivalent arsenic

MM A: Monomethylarsonic acid

DMA: Dimethylcin acid

TMAO: Trimethylarcinoxide

A B: Arsenobetaine (trimethylarsonium acetate)

DMAA: Dimethyl arsonium acetate

Me Co: Methylcobalamin

GSH: glutathione (reduced)

i S e (IV): Inorganic selenium (tetravalent) MIAA: Monoacetic acid

Example 1

[Reaction scheme]

Me C o

i A s (I I I) → MM A

G S H

[Reaction operation]

1. In a 5 ml Eppendorf tube, 740 L of reaction buffer (20 mM Tris_HGI (pH 7.6)) was added. To this, 22O U of a 10 OmM aqueous solution of GSH was added, and the mixture was stirred for 30 seconds at V o I t e x and allowed to stand at 37 ° C for 30 minutes. To this, 20 liters of 100 ppm inorganic arsenic (II I) standard solution (for atomic absorption) was added and stirred for 30 seconds. To this was added 20 L of a 7.4 mM methylcobalamin (MeCO) aqueous solution (Composition A). This was reacted in a constant temperature bath maintained at 37 ° C. and sampled periodically to track the increase in product.

[Analysis of product]

To compare the retention time of a standard sample with that of a reactive organism with an inductively coupled plasma ion mass spectrometer (Agilent 7 500 ce) connected directly and online with a high performance liquid chromatograph (Agilent 1100) We conducted more qualitative and quantitative analysis.

Example 2

[Reaction scheme]

Me C o

i A s (I I I) → MM A

G S H, i S e (I V)

[0101] [Reaction operation]

In Composition A of Example 1, 1 000 ppm of inorganic Se (IV) standard solution (atomic absorption The same procedure as in Example 1 was carried out except that 20 L of light was added (Composition B).

Comparative Example 1

The same procedure as in Example 1 was carried out except that MeCo was not added in Example 1 (Composition C). Table 3 shows the detoxification of inorganic arsenic to MMA (Example 1) and the detoxification of inorganic arsenic to DMA (Example 2).

[Table 3]

As shown in Examples 1 and 2, methylarsenic (MMA) was generated with the passage of time as compared to the comparative example. In Example 2, further methylation progressed, and formation of dimethyl arsenic (DMA) was also confirmed. In the presence of Me 2 Co, toxic inorganic arsenic has a remarkable effect of being detoxified to less toxic methylated arsenic.

[0105] In addition, the methylation of selenium was also examined. FIG. 31 shows the H P L C — I C P-MS chromatogram of the methylation reaction of hyposelene [Se (IV)] by MC. In the figure, A: Standard sample. B: sample after reaction, Se (IV): selenium acid, DMSe: dimethylselenium.

As shown in FIG. 31, it was confirmed that selenium acid was converted to less toxic dimethylselenium.

Example 3

[Reaction scheme]

Me C o

MMA → DMA

GSH [Reaction operation]

The same procedure as in Example 2 was carried out except that 20 L of MMA of 1000 ppm was added to the composition B of Example 2 (Composition D).

Comparative Example 2

Example 3 was carried out in the same manner as Example 3, except that MeGo was not added.

Example 4

[Reaction scheme]

Me C o

DMA → TMAO

G S H

[Reaction operation]

The same procedure as in Example 2 was carried out except that 20 L of 100 ppm p DMA was added to the composition B of Example 2 (Composition E).

[01 12] Comparative example 3

Example 4 was carried out in the same manner as Example 4 except that MeGo was not added. Table 4 shows the detoxification of MMA to DMA (Example 3) and the detoxification of DMA to TMA 0 (Example 4).

[0113]

Ho 4]

As shown in Example 3 _ !! to 3-3, the concentration of dimethyl arsenic (DMA) increased with the passage of time. In Comparative Example 2, generation of DMA was not observed. As shown in 4_1 to 4_3, it was found that the concentration of trimethylated arsenic (TMA0) was increased, and the arsenic substrate was converted to the most harmless trimethylated arsenic. The formation of trimethylated arsenic was not observed.

Example 5

[Reaction scheme]

MeCo, MIAA

TMAO → AB

G S H

[Reaction operation]

A composition B was prepared in the same manner as in Example 2 except that 20 L of 1 000 p p m TMAO was added instead of the inorganic arsenic in the composition B of Example 2 (Composition F)

Example 6

[Reaction scheme]

MIAA TMAO → AB

G S H

[Reaction operation]

A composition F was prepared in the same manner as in Example 5 except that MeGo was not added. (Composition G)

Example 7

[Reaction scheme]

MeGo MIAA

DMA → AB

G S H

[Reaction operation]

A composition F was prepared in the same manner as in Example 5 except that DMA was used instead of TMA0 (composition G). Table 5 shows the conversion to AB.

[Table 5]

[0122] As shown in Example 5, conversion of the arsenic substrate TMAO to AB was confirmed in the presence of MeGo and MIAA. As shown in Example 6, conversion of TMAO to AB was confirmed only in the presence of MIAA. As shown in Example 7, conversion of the arsenic substrate DMA to AB was confirmed in the presence of MeGo and MIAA. Example 8

First, the abbreviations used in the following examples will be described as follows.

i A s (I I I): Inorganic trivalent arsenic,

MM A: Monomethylarsonic acid

DMA: Dimethylcin acid

TMAO: Trimethylarcinoxide

A B: Arsenobetaine (trimethylarsonium acetate)

DMAA: Dimethyl arsonium acetate

Me Co: Methylcobalamin

GSH: glutathione (reduced)

M I A A: Monoacetic acid

AS: Arseno Sugar

i S e (IV): Inorganic selenium (tetravalent)

[0124] (1)

Microalgal culture

The microalga Chlorella (Ghlorel la vulgaris I AMG-629 strain) precultured to the logarithmic growth phase is inoculated in 150 ml of Bold's Basal (BB) Medium to a density of 1 x 10 ⁶ cel Is / ml, Stationary culture was carried out at a temperature of 25 ° C. under fluorescent lamp irradiation (4000 Lux, 24 hr irradiation). At this time, the medium was adjusted to the culture medium to which 1 OmM glucose or 1 OmM sodium acetate was added as a carbon source.

(2)

Arsenic uptake test

Arsenic acid was added so that it became 1 ppm as metal arsenic after inoculation, and the uptake | capture test of arsenic was implemented by culture | cultivating for 284 hours after arsenic addition.

(3)

Arsenic content measurement Inorganic arsenic and organic arsenic in algal cells can be detected with an inductively coupled plasma mass spectrometer (Agilent 7500 ce) directly connected online with a high performance liquid chromatograph (Agilent 1 100) as a standard sample, Qualitative and quantitative analysis was performed by comparing the retention times of the responding organisms.

[0127] (4)

Analysis conditions

As standard samples of organic arsenic compounds, MMA, DMA, TMAO, TeΜΑ, ΑΒ, AC are reagents of trichemical laboratory, and standard samples of inorganic arsenic are As (III), As (V) The sodium salt of Wako Pure Chemical's special grade reagent was used. The 100 mg / 100 mL standard solution of each arsenic compound was prepared by diluting with ultrapure water (Millipore).

[0128] I CP_MS device condition>

RF forward power: 1. 6 kW

R F reflect power: <1 W

Carrier gas flow: Ar 0.75 L / min

Samp I i ng 8.5 mm

Monitoring mass: m / z = 75 and

35 internal standard m / Z = 71

Dwel l time: 0.5 sec 0.01 sec

0.1 sec

Times of scan: 1 time

H P L C condition

Eluent: 5mM Nitric Acid / 6mM Ammonium Nitrate / 1.5mM Pyridinedicarboxylic Acid

Eluent flow rate: 0.4 m L / min

Injection volume: 20 L

Column: cation exchange column Shodex RSpak NN-414 (150 countries x 4.6 countries d.) Column temperature: 40 degrees Extraction of Arsenic Compounds from Microalgae Incorporating Arsenic

A microalgal extract (chlorella extracted with methanol and the methanol removed by evaporation) was prepared. The pure water was added to this and diluted, and the solution of the density | concentration of the following table was obtained. The components of UN 1 (Unknown "!) And UN 6 were assigned as compounds corresponding to arsenosugar (Fig. 1). Fig. 1 shows the HPL C-ICP-MS analysis of the extract of Quailella. (Upper: standard sample, lower: sample) Table 6 shows the concentration of arsenic compound (ppm) in the chlorella extract.

[Table 6]

A s (y) A 3 (III) MMA DMA UN 1 UN 6 Total Arsenic

2, 65. 0, 175 0, 62 0.035 1.1 1.22 S. 83

[Conversion to AB]

Reaction buffer (20 mM Tris_HGI (pH 7.6)) 7 40 1 _ was added to a volume of 1.5 ml Eppendorf tubes. To this was added 200 U of a 2OmM aqueous solution of GSH, and the mixture was stirred for 30 seconds at VoItex and allowed to stand at 37 ° C for 30 minutes. 100 L of the chlorella extract was added and stirred for 30 seconds. To this was added 135 liters of a 7.4 mM methylcobalamin (MeCo) aqueous solution. To this, 68 mg (0.35 / J M) of M AA was added and dissolved. This was reacted in a constant temperature bath maintained at 37 ° C., and periodically sampled to track the amount of increase in the product. As shown in FIG. 2, generation of AB could be confirmed when GSH, MeCo and MIAA were present. Figure 2 shows HPL C_ ICP-MS analysis of Chlorella extract (upper: standard sample, middle: GSH added (NEJ4-7), lower: MeCo + GSH + MI AA added (NEJ5-7)) FIG.

Example 9

Al force Lee Me CO, M I AA

A S → DMA → A B

G S H

[0133] [Conversion of Arsenosugar to DMA] 100 L of the chlorella extract was mixed with 1 N aqueous solution of 4 N a OH (1 mL) and allowed to stand at 80 ° C. It was confirmed that DMA was generated and that alsenosugars were converted to DMA (Fig. 3).

[Conversion from DMA to A B]

It carried out by carrying out similarly to the thing as described in Examples 1-7. FIG. 3 shows Chlorella extract to which GSH was added (NEJ4-4), GSH + MeCO + MIAA addition (NEJ5-4) to which NaOH treatment (bottom). Figure 4 also shows the addition of G S H + M e C o + M I A A to DMA (NE-9-4).

Example 1 0

Furthermore, experiments were conducted using methylcobalamin as a cobalt complex. First, 10 mg of methylcobalamin (Wako Pure Chemical Industries, Ltd.) was collected in a 1.5 mL Eppendorf tube. To this, 1 mL of ultrapure water (18 MQ / cm) was added to dissolve methyl cobalamin (7.4 country ₀ //) (solution A). 30.7 mg of glutathione (reduced form) was placed in a 1.5 mL Eppendorf tube, and dissolved in 1 mL of ultrapure water (100 liters / L) (solution B). Prepared an arsenous acid aqueous solution (for atomic absorption: 100 ppm: as metal arsenic)

(Solution G). An aqueous solution of selenite (for atomic absorption: 1000 ppm: as metal selenium) was prepared (solution D). A 100 mmol / L Tris-HGI buffer solution was prepared (pH 7.8, pH was adjusted with 0.01 mol / L hydrochloric acid aqueous solution) (solution E). The solution E was added 720 and the solution G was added 20 into a 1.5 mL Eppendorf tube, and the solution B was added 220 and allowed to stand at 37 ° C. for 1 hour. The solution A was added to the solution A and the solution D was added to the solution D, and the mixture was allowed to react in a 37 ° C. constant temperature bath. The reaction conditions are as follows.

Substrate concentration: [As] = 30 ^ mol / L

Natural vitamin B12 (methyl cobalamin) concentration: [MeGo] = 150; umol / L

Glutathione (reduced) concentration: [GSH] = 22 countries ol / L

Selenium concentration: [Se] = 760 卿 I / L

Buffer solution: 100 mM Tris-HGI buffer solution (pH 7.8), reaction temperature: 37 ° C., reaction solution: pH 3 [0137] — 50 1_ is sampled at fixed time intervals, diluted 10 fold with ultra pure water And HPLG Qualitative and quantitative analysis was performed by the -IGP-MS method (No. 8 in Table 7)

[Table 7]

FIG. 7 shows a time-dependent change of the concentration of the arsenic compound in the reaction solution (the results of Table 7 are graphed). In addition, 50 1_ was sampled from the reaction solution, which was treated with an aqueous solution of hydrogen peroxide (37 ° C. for 1 hour), diluted 10 times with ultrapure water, and the reaction product was similarly analyzed. (No. 9 1 1 in Table 7) The HPLG-IGP-MS chromatogram is shown in Figure 5 and Figure 6.

Example 1 1

Example 10 was carried out in the same manner as Example 10 except that the solution B was adjusted to pH 7 in 100 mM Tris_HGI buffer solution.

The results are shown in Table 8 and FIGS. 8 to 17. About 50% of the arsenous acid was methylated. Table 8 shows the concentrations of arsenic compounds in the reaction solution.

[0142] Ho 8]

* No. 1 7.7: before Η ₂ 0 ₂

Wood trees No. 8 Ν. 14: after H ₂ 0 ₂ treatment.

As apparent from Table 8, it was found that it is possible to perform the detoxification treatment by setting the number of oxidations to As (lll) → As (V) by the H ₂ 0 ₂ treatment to a high oxidation number. did.

Example 1 2

The procedure of Example 10 was repeated except that the pH of the reaction solution after preparation was changed to the value shown in Table 9 in Example 10.

The results are shown in Table 9 and FIGS. 18 to 29. For pH 6.7, about 50% of the arsenous acid was methylated. On the other hand, for pH 9, methylation did not proceed. Table 9 shows the concentrations of arsenic compounds in the reaction solution.

[0146]

Ho 9]

Example 1 3

Next, we tried the synthesis of [Gob (ll)] GI0 ₄ from Shianokobaramin. 1. Oxidation of cobalt complex ■ Reduction, methylation

(1)

Synthesis of [Gob (ll)] GI 0 ₄ from cyanokobalamin

[Reaction scheme 1]

[Formula 1]

[Experimental Operation]

Dissolve 50 mg of cyanocobalamin (Gob (I II) in [Chemical formula 1]) in 100 mL of methanol and degas by nitrogen bubbling. Add 400 mg (1.05 mol) of NaBH ₄ and check the green color from Co (l). Add 3 mL of 60 ° / o _HCI 0 _4ap . Add 50 mL of water and extract with methylene chloride. After washing with water, it was dried over anhydrous sodium sulfate and evaporated to dryness. Reprecipitation with benzene / n-hexane gives an orange powder.

(2)

Reduction of [Cob (l I)] to [Gob (l)]

[Reaction scheme 2]

[Chemical Formula 2]

Cob (ll) Cob (l)

[0152] [Experimental Procedure] [Gob (ll)] to GI0 ₄ 30 mg was dissolved in methanol 100 mL, degassed with nitrogen Baburi ring. Add 300 mg (0.788 mol) of NaBH ₄ and check the green color of Go (I). The cobalt complex (III) in the alkylated composition of the present invention can be converted to cobalt complex (II) from the reaction from Gob (II) to Gob (l) as shown in Reaction Scheme 2 The cobalt complex (II) thus obtained can be reduced with a photocatalyst or with a chemical reducing agent as shown in Example 14 below to obtain a cobalt (I) complex. The cobalt (I) complex can also be used as a substrate for a dehalogenation reaction. That is, it is possible to make the organic halogen compound harmless by using the cobalt (I) complex thus obtained.

Example 1 4

Next, we investigated the synthesis of [MeGob (ll)] by the reaction of [Gob (l)] with GH ₃ I (dehalogenation reaction of halide).

(3) Synthesis of [MeGob (ll)] by the reaction of [Gob (l)] and GH ₃ I (dehalogenation reaction of halide) [Reaction scheme 3]

[Formula 3]

X ⁼ CH3

Cob (l)

Cob (ll)

[Experimental Operation]

GH ₃ I 37 mg of (2.6x10- ⁴ mol) was added under the dark and stirred for 5 minutes.

Thus, the dehalogenation reaction occurs with the cobalt complex and the organic halogen compound. That is, not only is the organohalogen compound which is a harmful compound harmlessized by dehalogenation, but the obtained cobalt (III) complex detoxifies harmful compounds such as arsenic by methylation. It can be a convenient substrate.

(4) Synthesis of [MeCob (l I)] to [MeGob (l I I)]

[Reaction scheme 4]

[Formula 4]

X = CH ₃ X = CH,

Cobi ") Cob (IM)

[Experimental Operation]

2 mL of 60 ° / o HCI _04ap was added. Add 50 mL of water and extract with methylene chloride. After washing with water, dry over anhydrous sodium sulfate and evaporate to dryness. Benzene / n-hexane Precipitation is carried out to obtain orange powder methyl cobalamin.

Thus, by utilizing the oxidation state of the cobalt complex in the alkylated composition of the present invention, it is possible to detoxify organic halogen compounds and to detoxify harmful compounds such as arsenic. That is, using the cobalt (I 11) complex obtained by the dehalogenation reaction between the cobalt (I) complex and the organic halogen compound, the cobalt (III) complex and the harmful compound (including arsenic and the like) can be used. By the reaction, the harmful compound can be methylated to render it harmless.

On the other hand, when the resulting cobalt (II) complex is reduced through any reaction, a cobalt (I) complex can be obtained, which in turn can be used again for the detoxification of the organohalogen compound. .

Example 1 5

Next, under certain conditions, the use of GSH and methylcobalamin was examined for the most efficient conversion to TM AO.

[0163] First, GSH (60 mg, 0.195 country oL), methyl cobalamin (MC) 10 mg (7.44 mg), Tris-HGI buffer solution (pH 8, OuD) were placed in a 0.2 mL Eppendorf tube. A standard solution (for atomic absorption, 100 ppm as arsenic) was added in 21_, placed on an aluminum block heater heated to 125 ° C., and allowed to react for a predetermined period of time. It was diluted by 30 to 30 times and analyzed by HPLG-IGP-MS The results are shown in Table 10 and Table 11.

10]

[Table 11]

Tables 10 and 11 show the results of HPLG-IGP-MS analysis when G S H concentration, arsenic concentration, temperature, etc. were changed. Table 10 shows concentration and Table 11 shows percentage.

[0165] As a result, under the conditions of this example, it is possible that MG115 in Table 10 and Table 11 can change to approximately 100% TMA0 with GSH with the best data. Yes.

Example 1 6

Next, we investigated the case of using cysteine (C ys) instead of GSH. First, 0.2 gm of Eppendorf tube as a reducing agent (Gys) (20 mg, 0.165 oL), methylcobalamin (MG) (20 mg, 14. oL), phosphate buffer (pH 6) instead of GSH. , 100 ^ L). Arsenic standard solution (for atomic absorption, 100 ppm as arsenic) was added to this, and it was placed on an aluminum block heater heated to 110 ° C. and reacted for a predetermined time. The reaction product was diluted 10 to 30 times with 10% aqueous hydrogen peroxide and analyzed by HPLG-IGP-MS. The results are shown in Table 12. 12]

[0167] Table 12 shows the results of HPLG-IG P-MS analysis when changing the concentration of cysteine, the concentration of arsenic, the temperature, and the like. As with GSH, it can be seen that good results can be obtained even with the use of cystine. In particular, looking at the ratio of TMAO, MG157-3P in Table 12 is a good result.

Example 1 7

Next, we also examined the case where homocysteine was used instead of GSH.

First of all, homocysteine (HGys) (5 mg, 16.3 ^ moL), methylcobalamin (MG) (20 mg, 14.9 UmoL), phosphate buffer (pH 6, I put 100 ^ L). Arsenic standard solution (for atomic absorption, 100 ppm as arsenic) was added to this, and it was placed on an aluminum block heater heated to 120 ° C. and reacted for a predetermined time. The reaction product was diluted 10 to 30 times with 10% aqueous hydrogen peroxide and analyzed by HPLG-IGP-MS. The results are shown in Table 13.

[0169] 13]

Table 13 shows the results of HPL G-IGP-MS analysis when changing the homocysteine concentration, the arsenic concentration, the temperature, and the like. As with GSH, it can be seen that good results are obtained even with homocysteine. In particular, focusing on the ratio of TMAO, MG163-2P in Table 13 is a good result.

Example 1 8

Next, in addition to GSH, it investigated about the case where a thioglycol was added. That is, the addition effect of the high boiling point solvent was examined. Specifically, thioglycol having an SH group (TG, HSCH 2 CH 20 H, boiling point: 157 ° C.), and dimethyl sulfoxide having no SH group (DMS 0, (GH 3) 2 S 0, boiling point: 189 ° C.) were used. .

First, GSH (4 mg, 13 ^ moL), methyl cobalamin (MG) (1 mg, 0.74 ^ moL), TG (5 L), Tris-HGI buffer (0.2 mg Eppendorf tube as a reducing agent) pH 8 (ul) was added. Arsenic standard solution (10 ppm as arsenic) was added to this, placed on an aluminum block heater heated to 120 ° C, and reacted for a predetermined time. The reaction product is diluted 10 to 30 times with 10% hydrogen peroxide solution, and the reaction product is diluted by HPLG-IGP-MS. Analyzed (the description of MG179 in Table 14 and Table 15)

Also, for MG 180 in Tables 14 and 15, 0.2 mL of Eppendorf tube as a reducing agent GSH (4 mg 13 ^ mo L), methylcobalamin (MG) (1 mg 0.74 L) DMS 0 5; uL), Tris-HGI buffer (pH 8 uD was added. Add 21_ of arsenic standard solution (10 ppm as arsenic) to this, and place on an aluminum block heater heated to 120 ° C, for a predetermined time The reaction product was diluted 10 30 times with 10% aqueous hydrogen peroxide and analyzed by HPLG-IGP-MS The results are shown in Tables 14 and 15.

[Table 14]

[Table 15]

Tables 14 and 15 show the results of HPLG-IGP-MS analysis using TG, DMSO in addition to GSH (in Table 14 and Table 15, GSH: glutathione ( Reduced form), MC: methylcobalamin, TG: thioglycol, DMSO: dimethyl sulfoxide). Table 14 shows the concentration, and Table 15 shows the percentage. As in the case of using GSH alone, it can be seen that good results can be obtained using TG DMSO.

Example 1 9

Next, the case where cysteine (C ys) was used as a reducing agent was further examined according to the above-mentioned example. Table 16 shows the methylation reaction of arsenite [iAs (lll)] by MC (acid condition) (reactant). [Table 16]

Table 17 shows the methylation reaction of arsenite [iAs (l l l)] by MC (acid conditions) (reaction conditions and reaction products). In the table, GSH: reduced glutathione, MA: methylcobalamin, As: starting material [here, trivalent inorganic arsenic: iAs (lll)], MMA: monomethylarsenic, DMA: dimethylarsuoxide, TMA O : Trimethylarcinoxide, TeMA: Tetramethylarsenic, conversion rate = 100% ([iAs (V)] + [MMA] + [DMA] + [TMA0] + [TeMA]) / [iAs Calculated as (lll)].

Ho 17]

Table 18 shows the methylation reaction of arsenous acid [iAs (l l l)] with MC (neutral conditions).

[Table 18]

Anti-JS substance solvent stoichiometry

3⁄4: Test Cys MC buffer (pH 8) Returning agent Methylating agent

(U mol) (WL) [Cys] / [As] [MC] / [As]

1 165.1 149 2.7 50 61902 5579

2 165.1 22.3 2.7 50 61902 8368

3 165.1 29.8 2.7 50 61902 11158

4 165.1 37.2 2.7 50 61902 13947

5 165.1 44.6 2.7 50 61902 16736

6 165.1 52.1 2.7 50 61902 19526

7 165.1 37.2 2.7 50 61902 13947 Table 19 shows the methylation reaction of arsenite [iAs (lll)] by MC (neutral conditions) (reaction conditions and reaction products).

Ho 19]

Table 20 shows the methylation reaction of arsenous acid [iAs (l l l)] by MC (alkaline condition) (reactant).

10]

Table 21 shows the methylation reaction of arsenate [iAs (lll)] by MC (alkaline conditions) (reaction conditions and reaction products). Ho 21]

Example 20

Next, the detoxification of antimony was similarly tested in accordance with the above-described example. FIG. 32 shows the HPLG-IGP-MS chromatogram (measuring element: Sb, m / z 121). In the figure, A: standard sample [iSb (l I l)], B: sample after reaction (MC + GSH), C: sample after reaction (MC only). Table 22 shows the methylation reaction (reactant and reaction conditions) of inorganic anthimone.

22]

[0183] Table 23 shows the methylation reactions (reaction products) of inorganic antimony 23]

As apparent from FIG. 32, it was found that by reacting methylcobalamin with trivalent inorganic antimony, U 1, U 2, U 3 and U 4 belonging to methylated antimony were formed. . Therefore, it is understood that it is possible to obtain methylated antimony which is more harmful to antimony.

Example 2 1

Next, it investigated about the influence which the molar ratio of each component of the composition for alkylation gives. Specifically, the experiment was conducted according to the above-described example using GSH as a reducing agent, arsenic as a harmful compound, and methylcobalamin as a cobalt complex. The results are shown in Table 24 and Table 25. Table 24 shows the methylation reaction (reactant) of arsenous acid [iAs (lll)] by MC. Table 25 shows the methylation reaction (reaction conditions and reaction products) of arsenite [iAs (lll)] by MC. In the table, [GS 2 H]: molar concentration of reducing agent (GS 2 H), [MC]: molar concentration of methylcobalamin, [A s]: molar concentration of arsenous acid which is a starting material. Further, in the table, GSH: reduced glutathione, MC: methylcobalamin, As: starting material [here, trivalent inorganic arsenic: iAs (lll)], MMA: monomethylarsenic, DMA: dimethyarsinho Texa, TMAO: Trimethyl alanoxide, T eMA: A tetramethylarsenic, conversion rate = 100% ([i As (V)] + [MMA] + [DMA] + [TMAO] + [T eMA]) Z Calculated as [iAs (l II)]. Also, FIG. 33 shows reaction conditions for formation of trimethylarsenic (TMA) from arsenous acid by methylcobalamin. Ho 24]

Ho 25]

As shown in Tables 24 and 25, when the reducing agent GSH is added in a 10000-fold excess with respect to arsenic, and when methylcobalamin is added with a excess of 1 000-fold excess with respect to arsenic, it is non-toxic Trimethylarcinoxide could be obtained at a relative ratio of 90% or more. That is, [GS H] / [A s]> 1 000, [MC] / [A s]> 100, more preferably, [GSH] / [A s]> 1 0000, [MC] / [A s] It was found that non-toxic trimethylarsoxide can be obtained in relative proportions of 90% or more when> 1 000.

Industrial applicability

The composition of the present invention can more practically and industrially provide a method for detoxifying harmful compounds that can contribute to the detoxification of harmful compounds including arsenic and the like. Hazardous compounds such as alkylated arsenic are converted to more harmless compounds, and the harmless compounds are extremely stable and safe, so it is widely used in areas such as industrial waste treatment, environmental protection of sludge, and soil. Is extremely effective in the field of

Claims

The scope of the claims

[I] A composition for alkylation containing a cobalt complex.

[2] The composition according to claim 1, characterized in that the harmful compound containing at least one element selected from the group consisting of arsenic, antimony and selenium is alkylated by using the above-mentioned cobalt complex. .

[3] Furthermore, at least one selected from the group consisting of arsenic, antimony and selenium

The composition according to claim 1, comprising a reducing agent that reduces one metal.

[4] The composition according to claim 3, wherein the reducing agent is a substance having a SH group.

[5] The substance having an SH group is at least one selected from the group consisting of glutathione, reduced glutathione (GSH), cysteine, S-adenosylcystine, sulforafuan, homocysteine, thioglycol, and the like. Composition as described.

[6] The composition according to any one of claims 1 to 5, further comprising a methylation additive having an S-Me group.

[7] The composition according to claim 6, wherein the methylation addition factor is at least one selected from the group consisting of methionine and S-adenosylmethionine.

[8] The composition according to any one of claims 1 to 7, further comprising a buffer.

[9] The composition according to claim 8, wherein the pH value of the buffer solution is in the range of 5 to 10.

[10] The composition according to any one of claims 1 to 9, wherein the pH of the alkylation composition is less than 9.

[II] The composition according to any one of claims 1 to 10, further comprising H ₂ O ₂

[12] The composition according to any one of claims 1 to 11, further comprising an organic halogen compound.

[13] The composition according to claim 12, wherein the organic halogen compound is methyl halide.

[14] The composition according to claim 13, wherein the methyl halide is at least one selected from the group consisting of methyl iodide, methyl bromide and methyl chloride.

15. The composition according to claim 12, wherein the organic halogen compound is a halogenated acetic acid.

[16] The composition according to claim 15, wherein the halogenated acetic acid is at least one selected from the group consisting of crocodile acetic acid, bromoacetic acid, and joud acetic acid.

[1 7] The organic halogen compounds are methyl chloride, methyl bromide, methyl iodide, chloroacetic acid, bromoacetic acid, bromoacetic acid, chloroacetic acid, chloroethanol, bromoethanol, bromoethanol, chloropropanoic acid The composition according to claim 12, wherein the composition is at least one selected from the group consisting of: bromopropionic acid, sodium propionic acid, cloroacetic acid ethyl ester, bromoacetic acid ethyl ester, sodium acetate ethyl ester.

[18] The organic halogen compound is represented by Chemical Formula 1, Chemical Formula 1 RMgX

A Grignard reagent represented by (wherein R = Me, GH ₂ G00H, or GH ₂ G00 G ₂ H ₅ in the chemical formula 1, and X = G and Br or I). 1 4 Composition described.

[19] The organohalogen compound is derived from a refractory organic substance selected from the group consisting of pesticides, flame retardants, dioxins, PGBs, DDTs, trihalomethanes, trichloroethols and croforms. 18. The composition according to any one of 18.

[20] The composition according to any one of claims 1 to 19, further comprising a reducing agent that reduces a cobalt complex.

[21] The composition according to claim 20, wherein the reducing agent is at least one selected from the group consisting of titanium oxide or ruthenium complex.

[22] The cobalt complex is methylcobalamin (methylated vitamin B12, formal name: Go HI [5], 6-dimethylbenz 1 1 H-imidazo 1 1 1 l 1 Go; 3− Methylcobamide], Vitamin B 12 such as Sanocobalamin, Cobalt (II) Acetylacetonate, Cobalt (III) Acetylacetonate, Cobaltocarponyl (Nicobalt Octacalponyl), Cobalt (| |) 1, 1, 1, 5, 5 , 5_ Hexafluoroacetylacetonato, Cobalt (II) Mesotetraphenylphenyl Porphyrin, Bis (pentamethylcyclopentagenyl) cobalt Hexafluorophosphate, Ν, Ν '-Bis (salicylidene) ethylenediamine cobalt ( II), bis (2, 2, 6, 6- tetramethyl_3, 5 _ heptanedionate) ko / lireto (11), (black mouth phthalocyaninato) ko / lireto (II), chloro tris (trifenyl phosphine) Cobalt (I), Methyl complex of cobalt (II) acetate, Cobalt (II) benzoate, Cobalt (II) cyanide, Cobalt (II) cyclohexanebutyric acid, Cobalt (II) 2-ethylhexylate, meso —Tetramethyoxyphenyl porphyrin cobalt (II), cobalt naphthenate, cobalt phthalocyanin cobalt (11), methyl cobalt (III ) Protoporphyrin IX, cobalt stearate, cobalt sulfamate (II), (1R, 2R) _ (-)-, 2-cyclohexanediamino-, Ν '-bis (3, 5-di _ t _ _ _ pyl) Salicylidene) cobalt

(II), (1S, 2S) _ (+) _1, 2-cyclohexanediamino-Ν, Ν '-bis (3, 5-di _ t _ pyl salicylidene) cobalt (II), cyclopentajenyl bis ( Trifenylphosphine) cobalt (I), cyclopentajenylcobalt dibasic sulfonyl, dibromobis (triphenyl phosphine) cobalt (||), (tetraaminochlorophthalocyaninato) co / lyreto (II), (tetra (Peptyl phthalocyaninato) A methyl complex of at least one compound selected from cobalt (II), or a cobalt monomethyl complex formed by coexisting the cobalt compound with an alkyl halide, particularly methyl halide. The composition according to any one of claims 1 to 21, which is at least one selected from the group consisting of

[23] The molar concentration of the reducing agent for reducing at least one metal selected from the group consisting of arsenic, antimony and selenium [Reducing Agent], and the molar concentration of a metal selected from arsenic, antimony and selenium [ The composition according to any one of claims 2 to 22, wherein the ratio to [Metal], that is, [Reduci ng Agent] / [Metal] is 1 000 or more.

[24] The composition according to claim 23, wherein the ratio is 10000 or more.

[25] The ratio of the molar concentration of the cobalt complex [Go comp l ex] to the molar concentration of a metal selected from arsenic, antimony, selenium [Metal], ie, [Go comp l ex] / [Metal l] The composition according to any one of claims 1 to 24, wherein the force is 100 or more.

[26] The composition according to claim 25, wherein the ratio is 1000 or more.

[27] A harmful compound containing at least one element selected from the group consisting of arsenic, antimony and selenium in the presence of the composition according to any one of items 1 to 26, and the composition according to item 1, The detoxification method of the said harmful compound detoxified by alkylating.

[28] The method according to claim 27, wherein the detoxification is carried out by setting the valence of said one element to a high oxidation number.

[29] The method according to claim 27, wherein at least one bond of the one element is alkylated.

[30] The method according to any one of claims 27 to 29, wherein the element is arsenic.

[31] The method according to any one of claims 27 to 30, wherein the 50% lethal dose (LD _M ) of the compound detoxified by the alkylation is at least 1000 mg / kg.

[32] The 50% cell growth inhibitory concentration (IG ₅₀ ) power of the compound detoxified by the alkylation is 1000; u M or more, any one of claims 7 to 31, and any one of claims 1 to 3. the method of.

[33] The harmful compound is selected from the group consisting of arsenous acid, arsenic pentoxide, arsenic trichloride, arsenic pentachloride, arsenic sulfide compounds, cyano-arsenic compounds, nitrogen-containing arsenic compounds, and other arsenic inorganic salts The method according to any one of claims 2 to 3.

[34] The method according to any one of claims 27 to 33, wherein the alkylation is methylation.

[35] The method according to claim 34, wherein the harmful compound is a dimethyl compound or a trimethyl compound by the methylation.

[36] The method according to claim 35, wherein the dimethyl compound is dimethylarsonylethanol (DMAE), dimethylarsonyl acetate (DMAA), dimethylarsinic acid, or arsenochegar.

[37] The method according to claim 35, wherein the trimethyl compound is arsenocholine, arsenobetaine, trimethylarsenosugar or trimethylarsinoxide.

[38] In the presence of the composition according to any one of claims 1 to 26, it is selected from the group consisting of pesticides, flame retardants, dioxins, PGB, DDT, trihalomethanes, trichloroethyls, oral forms. The detoxifying method of organic halides which detoxifies the organic halides by dehalogenating.

[39] A compound selected from the group consisting of pesticides, flame retardants, dioxins, PGBs, DDTs, trihalomethanes, trichloroethyls, oral forms in the presence of the composition according to any one of claims 1 to 26. The organic halide is detoxified by dehalogenation to contain at least one element selected from the group consisting of arsenic, antimony and selenium in the presence of the cobalt complex obtained by the reaction. The method according to any one of claims 26 to 37, wherein the harmful compound is detoxified by alkylation.

[40] The method according to any one of claims 26 to 37, further comprising: light irradiation in the presence of a reducing agent that reduces the cobalt complex.

[41] The method according to claim 40, wherein the reducing agent is at least one selected from the group consisting of titanium oxide or ruthenium complex.