WO2008012935A1 - Alkylation composition, and method for detoxification of toxic compound using the composition - Google Patents

Abstract

Disclosed are: an alkylation composition which is useful for detoxification of inorganic arsenic or the like; and a safe and highly efficient detoxification method using the composition. The alkylation composition comprises an organic metal complex having a cobalt-carbon bond. The detoxification method is characterized by detoxifying a toxic compound having at least one element selected from the group consisting of arsenic, antimony and selenium by alkylating the toxic compound in the presence of the compound.

Description

 Specification

 Composition for alkylation and method for detoxifying harmful compounds using the composition

 Technical field

 TECHNICAL FIELD [0001] The present invention relates to an alkylating composition and a method for detoxifying harmful compounds using the composition.

 Background art

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

 Conventionally, as a method for removing these heavy metals, a flocculant such as polyaluminum chloride (PAC) is added to waste water containing inorganic arsenic such as toxic arsenous acid, and arsenic is added to the iron in the flocculant and raw water. A method of agglomerating, adsorbing, precipitating, and removing by filtration, or a method of adsorbing an arsenic compound with activated alumina or a cerium-based adsorbent is generally known.

 [0004] On the other hand, in the natural world, it is clear that marine organisms such as seaweed accumulate inorganic arsenic, and that part of the inorganic arsenic is converted to organic arsenic compounds such as dimethylated arsenic by physiological reactions. (Non-Patent Document 1). These organic arsenic compounds are generally known to exhibit lower toxicity to mammals than inorganic arsenic. In particular, arsenic contained in seafood is mostly present as arsenobetaine. This arsenobetaine is internationally recognized as non-toxic arsenic.

 [0005] Non-Patent Document 1: Kai seet a l., 1998, Organomet. Chem.. 12 137-143

 Disclosure of the invention

 Problems to be solved by the invention

[0006] However, in the above-described method of removing heavy metals using filtration, adsorption, etc., sludge containing harmful compounds such as inorganic arsenic that remains harmful, and It is necessary to store or landfill the adsorbent on which the harmful compound is adsorbed after sealing it with concrete etc. so that the harmful compound does not leak to the outside. Therefore, there is a problem that a large amount of processing is difficult.

 [0007] Therefore, in order to solve the above-mentioned problems, the present invention provides a composition useful for efficiently and systematically detoxifying harmful compounds including arsenic and the like, and harmful using the composition An object is to provide a method for detoxifying a compound.

 Means for solving the problem

[0008] In order to achieve the above object, the present inventors methylated a harmful compound containing arsenic or the like by a chemical reaction using an organic metal complex having a cobalt-carbon bond, particularly dimethylated, more preferably Tried to trimethylate, and as a result of intensive studies on the methylation reaction of the harmful compound, the present invention was found.

[0009] That is, the alkylating composition of the present invention is an alkylating composition containing an organometallic complex having a cobalt single-carbon bond, and the organometallic complex is methylaquocobyrinic acid heptamethyl ester perchloric acid. Salt [(CH ₃ ) (H ₂ 0) Cob (l I l) 7C _ie ster] CI 0 ₄ [Chemical 1]

 [Chemical 1]

 [(CHsXHaOJCobilllJ C! EsterlCIOJ

 It is characterized by being.

Also, in a preferred embodiment of the alkylated composition of the present invention, the organic By using a metal complex, a harmful compound containing at least one element selected from the group consisting of arsenic, antimony, and selenium is alkylated.

 [001 1] Further, in a preferred embodiment of the alkylated composition of the present invention, the composition further comprises a reducing agent that reduces at least one metal selected from the group consisting of arsenic, antimony, and selenium. To do.

 [0012] In a preferred embodiment of the alkylation composition of the present invention, the reducing agent is a substance having an SH group.

[0013] Further, in a preferred embodiment of the alkylated composition of the present invention, the substance having an SH group is selected from the group consisting of dartathione, reduced dartathione (GSH), cysteine, S_adenosylcystine, and sulforan. It is characterized by at least one species.

[0014] In a preferred embodiment of the alkylated composition of the present invention, S

It contains a methylation additive having a -Me group.

[0015] In a preferred embodiment of the alkylation composition of the present invention, the methylation additive factor is at least one selected from the group consisting of methionine and S-adenosylmethionine. And

[0016] In a preferred embodiment of the alkylated composition of the present invention, the composition further comprises a buffer solution.

[0017] In a preferred embodiment of the alkylation composition of the present invention, the pH of the buffer solution is in the range of 5 to 10.

[0018] In a preferred embodiment of the alkylated composition of the present invention, the composition further comprises an organic halogen compound.

[0019] In a preferred embodiment of the alkylated composition of the present invention, the organic halogen compound is methyl halide.

[0020] In a preferred embodiment of the alkylated composition of the present invention, the halogenated methyl is at least one selected from the group consisting of methyl iodide, methyl bromide, and methyl chloride. And [0021] In a preferred embodiment of the alkylated composition of the present invention, the organic halogen compound is a halogenated acetic acid.

 [0022] In a preferred embodiment of the alkylated composition of the present invention, the halogenated acetic acid is at least one selected from the group consisting of cloacic acetic acid, bromoacetic acid, and sodoacetic acid. To do.

 [0023] Further, in a preferred embodiment of the alkylation composition of the present invention, the organic halogen compound is selected from the group consisting of methyl chloride, methyl bromide, methyl iodide, black mouth acetic acid, bromoacetic acid, sodoacetic acid, black mouth ethanol, bromo It is at least one selected from the group consisting of ethanol, chloroethanol, chloropropionic acid, bromopropionic acid, chloropropionic acid, chloroacetic acid ethyl ester, bromoacetic acid ethyl ester, and chloroacetic acid ethyl ester.

[0024] Further, the detoxification method of the present invention comprises at least 1 selected from the group consisting of arsenic, antimony, and selenium in the presence of the composition according to any one of claims 1 to 16. It is characterized by detoxifying harmful compounds containing seed elements by alkylation.

[0025] In a preferred embodiment of the detoxification method of the present invention, the detoxification method is characterized in that the valence of the one element is made high oxidation number.

[0026] In a preferred embodiment of the detoxifying method of the present invention, at least one bond of the one element is alkylated.

[0027] In a preferred embodiment of the detoxification method of the present invention, the element is arsenic.

[0028] In a preferred embodiment of the detoxification method of the present invention, the 50% lethal dose (LD ₅₀ ) of the compound detoxified by the alkylation is 1000 mg / kg or more. .

[0029] Further, in a preferred embodiment of the detoxification method of the present invention, the compound detoxified by the alkylation has a 50% cell growth inhibitory concentration (IC ₅₀ ) of 1000 M or more. To do. [0030] Further, in a preferred embodiment of the detoxification method of the present invention, the harmful compound is arsenous acid, arsenic pentoxide, arsenic trichloride, arsenic pentachloride, arsenic sulfide compound, cyanoarsenic compound, black arsenic compound It is selected from the group consisting of compounds, and other arsenic inorganic salts.

 [0031] Further, in a preferred embodiment of the detoxification method of the present invention, the alkylation is methylation.

[0032] Further, in a preferred embodiment of the detoxification method of the present invention, the harmful compound is a dimethyl compound or a trimethyl compound by the methylation.

 [0033] Further, in a preferred embodiment of the detoxification method of the present invention, the dimethyl compound is dimethylarsonyl ethanol (DMAE), dimethylarsonyl acetate (DMM), dimethylarsinic acid, or arseno sugar. It is characterized by being.

 [0034] Further, in a preferred embodiment of the detoxification method of the present invention, the lymethyl compound is characterized in that it is arsenocholine, arsenobetaine, trimethylarsenosuga or arlymethylarsinoxide.

 The invention's effect

 [0035] The composition for alkylation of the present invention has an advantageous effect that it can easily and easily alkylate harmful compounds, particularly harmful compounds containing arsenic, antimony, selenium and the like. In addition, according to the method of the present invention, harmful compounds can be rendered innocuous as much as possible, and there is an advantageous effect that a large space such as a storage place is not required. In addition, according to the method of the present invention, since the living object itself is not used as it is, an advantageous effect is achieved in that unnecessary by-products are not generated. Furthermore, according to the present invention, there is an advantageous effect that harmful inorganic arsenic can be reduced with a simple operation.

 Brief Description of Drawings

[0036] FIG 1 shows an electronic spectrum of Co (lll) complex of vitamin B _12. (solvent: Methylene chloride) A represents (CN) ₂ Cob (l II ester), B represents [(CN) (H ₂ 0) Cob (11 I ester! ICIiXt, respectively).

[FIG. 2] FIG. 2 shows the electronic spectrum of the Co (ll) complex of vitamin B ₁₂ (solvent: methyl chloride). A represents the case of [Cob (ll) 7C ₁ ester] CI0 ₄ (base-off type), and B represents the case of [Cob (II) 7dester] C 10 ₄ + pyridine (base-on type).

[FIG. 3] FIG. 3 shows the electronic spectrum of the Co complex of vitamin B ₁₂ (solvent: methylene chloride). In Fig. 3, A is [(CH ₃ ) (H ₂ 0) Cob (lll) 7C ₁ ester] CI0 ₄ (solvent: methylene chloride, before light irradiation). Each case is shown as a spectrum.

[Figure 4] Figure 4 shows the HP LC_ I CP-MS chromatogram of the product of the methylation reaction of inorganic arsenic with [(CH ₃ ) (H ₂ 0) Cob (111) 7C _ie ster] C 10 ₄ . (A) After reaction for 30 minutes, (B) After 4 hours of reaction.

 FIG. 5 shows an HPLC-ICP-MS chromatogram. (No. in the figure corresponds to No. in Table 3)

 FIG. 6 shows an HPLC-ICP-MS chromatogram. (No. in the figure corresponds to No. in Table 3)

 [FIG. 7] FIG. 7 shows the change over time in the concentration of the arsenic compound in the reaction solution (No. 1 to No. 8 in Table 3 are plotted).

 [FIG. 8] FIG. 8 shows the change over time in the ratio of arsenic compounds in the reaction solution (Nos. 1 to 7 in Table 3 are graphed).

 [FIG. 9] FIG. 9 shows the change over time in the ratio of the arsenic compound in the reaction solution (Nos. 6 to 11 in Table 3 are graphed).

 [FIG. 10] FIG. 10 shows an HPLC-ICP-MS chromatogram (corresponding to No. in Table 4).

[Figure 11] Figure 11 shows the HPLC-ICP-MS chromatogram (corresponding to No. in Table 4).

[Figure 12] Figure 12 shows the HPLC-ICP-MS chromatogram (hydrogen peroxide treatment) (corresponding to No. in Table 4).

[Figure 13] Figure 13 shows the HPLC-ICP-MS chromatogram (corresponding to No. 12 to No. 14 in Table 4). [FIG. 14] FIG. 14 shows the change with time in the concentration of the arsenic compound in the reaction solution (hydrogen peroxide solution untreated).

 [FIG. 15] FIG. 15 shows the change over time in the concentration of the arsenic compound in the reaction solution (after treatment with hydrogen peroxide solution).

 [FIG. 16] FIG. 16 shows the change over time in the ratio of arsenic compounds in the reaction solution (hydrogen peroxide water untreated).

 [FIG. 17] FIG. 17 shows the change over time in the ratio of arsenic compounds in the reaction solution (after treatment with hydrogen peroxide solution).

 [FIG. 18] FIG. 18 shows the change with time of the ratio of the arsenic compound in the reaction solution.

 FIG. 19 shows an HPLC-ICP-MS chromatogram.

 FIG. 20 shows an HPLC-ICP-MS chromatogram.

 BEST MODE FOR CARRYING OUT THE INVENTION

[0037] The alkylated composition of the present invention comprises methyl acocobilic acid heptamethyl ester perchlorate [(CH ₃ ) (H ₂ 0) Cob (l

I I ester! ICIiXt [Chemical 2]

[0038] [Chemical 2]

 [(CHsXHaOJCobilllJ C! EsterlCIOJ

 Containing.

That is, in the alkylating composition of the present invention, a harmful compound containing at least one element selected from the group consisting of arsenic, antimony, and selenium is alkylated by using the organometallic complex. Is possible . Here, in this specification, a harmful compound means a compound that may have some adverse effect on an organism when it is released into the environment and exposed to the organism.

[0040] Among the harmful compounds, arsenic-containing harmful compounds include arsenous acid, arsenic pentaoxide, arsenic trichloride, arsenic pentachloride, arsenic sulfide compounds, cyanoarsenic compounds, chloroarsenic compounds, and other Examples include arsenic inorganic salts. These arsenic, for example, has an LD _{5 ()} (mg / kg) (50% lethal dose in mice) of 20 or less, and is generally toxic to organisms.

 [0041] Examples of harmful compounds containing antimony include antimony trioxide, antimony pentoxide, antimony trichloride, and antimony pentachloride.

 [0042] Further, examples of harmful compounds containing selenium include selenium dioxide and selenium trioxide.

 [0043] 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. The presence of such a reducing agent can further promote alkylation. There is a possibility that the ability to reduce arsenic and the methyl transfer reaction may be rate-limiting in the conversion of arsenic to arsenobetaine, but it is thought that conversion to arsenobetaine etc. can be promoted by adding a reducing agent. It is done. Examples of such a reducing agent include substances having an SH group. Specifically, substances having an SH group include dartathione, reduced dartathione (GSH), cysteine, S-adenosylcysteine, Mention may be made of at least one selected from the group consisting of sulfaurafuan.

[0044] In a preferred embodiment of the alkylation composition of the present invention, it further contains a methylation additive having an S-Me group. If the methylation additive is present, more alkyl groups can be provided, and more alkylation and detoxification can be achieved. The methylation additive factor is at least one selected from the group consisting of methionine and S-adenosylmethionine. Can be mentioned.

 [0045] The alkylating composition of the present invention may contain a buffer. Buffer solution

In general, those used for isolation, purification, storage, etc. of biomaterials can be used, and are not particularly limited. For example, squirrel buffer solution, phosphate buffer solution, carbonate buffer solution, boric acid A buffer solution such as a buffer solution can be exemplified. In addition, the pH of the buffer solution is preferably in the range of 5 to 10 in consideration of safer detoxification.

 [0046] The alkylated composition of the present invention may further contain an organic halogen compound. From the viewpoint of facilitating the conversion of dimethyl compound and Z or trimethyl compound to arsenobetaine, examples of the organic halogen compound include halogenated methyl. Examples of the methyl halide include at least one selected from the group consisting of methyl iodide, methyl bromide, and methyl chloride from the viewpoint of high methylation reactivity.

 [0047] In addition, examples of the organic halogen compound include at least one selected from the group consisting of sodoacetic acid, sodoethanol, bromoacetic acid, bromoethanol, and sodopropionacetic acid from the viewpoint of high alkylation reactivity. .

 [0048] In a preferred embodiment, the organic halogen compound is a halogenated acetic acid. Examples of the halogenated acetic acid include at least one selected from the group consisting of black mouth acetic acid, bromoacetic acid, and odoacetic acid.

 [0049] In a preferred embodiment, the organic halogen compound includes methyl chloride, methyl bromide, methyl iodide, black mouth acetic acid, bromoacetic acid, sodoacetic acid, black mouth ethanol, bromoethanol, sodoethanol, chloro Examples thereof include at least one selected from the group consisting of oral propionic acid, bromopropionic acid, odopropionic acid, chloroacetic acid ethyl ester, bromoacetic acid ethyl ester, and chloroacetic acid ethyl ester.

[0050] Next, the detoxification method of the present invention will be described. That is, the method for detoxifying a harmful compound of the present invention comprises arsenic in the presence of the alkylating composition of the present invention described above. A detrimental compound containing at least one element selected from the group consisting of antimony and selenium is detoxified by alkylation. Here, the composition for alkylation and harmful compound of the present invention mean those described above, and can be applied as it is in the detoxification method of the present invention.

 [0051] In a preferred embodiment of the detoxification method of the present invention, 50% cell growth inhibitory concentration

From the viewpoint that (IC ₅₀ ) or LD ₅₀ is large and more detoxification can be achieved, detoxifying the harmful compound by increasing the valence of the above-mentioned one kind of element contained in the above hazardous compound Is preferred. Specifically, the above-described composition of the present invention can be used as a catalyst for the reaction, and the valence of the one kind of element can be increased to a high oxidation number by alkylation. In the case where the above element is arsenic or antimony, it is preferable that the trivalent valence is pentavalent, and in the case of selenium, the tetravalent valence is hexavalent.

 In the present invention, detoxification of the harmful compound is performed by alkylating the harmful compound. Here, detoxification can be achieved by alkylating at least one bond of the one kind of element in the harmful compound.

 [0053] Specifically, by performing the reaction using the above-described alkylating composition of the present invention, at least one bond of the one kind of element can be alkylated. Here, examples of the alkyl group added to the one kind of element include a methyl group, an ethyl group, and a propyl group. From the viewpoint of achieving detoxification more efficiently, a methyl group is preferred as the alkyl group.

In the detoxification method of the present invention, from the viewpoint of safety to the living body, 50% lethal dose (LD ₅₀ ) of the compound detoxified by the above alkylation (50% of mice die) The oral toxicity due to drug dose) is preferably 1000 mg / kg or more, more preferably 5000 mg / kg or more.

[0055] Further, in the detoxification method of the present invention, from the viewpoint of safety to the living body, the 50% cell growth inhibitory concentration (IC ₅₀ ) of the compound detoxified by alkylation or arylation is used. 1000 M or more, preferably 3000 M or more It is more preferable that As used herein, 50% cell growth inhibitory concentration (IC ₅₀ ) is a numerical value indicating the concentration of a substance necessary to inhibit or inhibit the growth of 100 cells with a substance by 50%. means. The smaller the IC _5Q value, the greater the cytotoxicity. IC ₅₀ was calculated from the results of examining the cytotoxicity of plasmid DNA damage under the condition of 37 ° C. for 24 hours.

Here, Table 1 shows IC ₅ o of each arsenic compound.

[0056] [Table 1]

* 50% Growth

 inhibition

 [0057] From Table 1, Arsenosugar with trivalent arsenic has higher cytotoxicity than monomethylated arsenic (MMA) and dimethylated arsenic (DMA), but with trivalent arsenic. It can be seen that the cytotoxicity is lower than that of MMA DMA having arsenic and arsenous acid. On the other hand, MMA DMA with trivalent arsenic has higher cytotoxicity than arsenous acid (trivalent and pentavalent), but overall, arsenic compounds with pentavalent arsenic are 3 from the viewpoint of cytotoxicity. It can be understood that the safety to living bodies is higher than that of arsenic compounds containing valent arsenic.

[0058] Table 2 shows LD ₅ Q of each arsenic compound. [Table 2]

* 5 0% Letha I

 dose

 [0059] In the detoxification method of the present invention, the biological half-life of the compound detoxified by the alkylation is preferably 8 hours or less from the viewpoint of safety to the living body. . In the detoxification method of the present invention, it is preferable that the harmful compound is 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 (DMM), dimethylarsinic acid, and arsenosugar. In addition, examples of the trimethyl compound include arsenocholine, arsenobetaine, “lymethylarsenosugar”, and “lymethylarsinoxide”.

 Example

 Examples of the present invention will be described below, but the following examples do not limit the scope of the present invention. First, abbreviations used in the examples will be described as follows.

 [0061] [abbreviation]

[(CH ₃ ) (H ₂ 0) Cob (1 1 1) 7dester] C 10 ₄ : Methyl acocobilic acid heptamethyl ester perchlorate

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

 M M A: Monomethylarsonic acid

DMA: Dimethylcinnic acid TMAO: Trimethylarsinoxide

 A B: Arsenobetaine (卜 -Limethylarsonium acetate)

 DM A A: Dimethylarsonium acetic acid

 AS: Arseno Sugar

 M e C o: Methylcobalamin

 GSH: Dartathion (reduced type)

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

M I A A: Monoodoacetic acid

 [Synthesis of cobalt complex]

[(CH ₃ ) (H ₂ 0) Cob (111) 7dester] C 10 ₄ : Synthesis of methylaquocobyrinic acid heptamethyl ester perchlorate

(1) Synthesis of (CN) ₂ Cob (l I l) 7dester

 [Reaction scheme]

[Chemical 3]

 The scheme of chemical formula 3 shows the reaction route from cyanocobalamin (left of chemical formula 3) to (CN Cobdll dester (right of chemical formula 3).

 [Experimental operation]

Shianokobaramin 1.0g of (7.5x10- ⁴ mol) was dissolved in methanol 300 mL, after cooling the concentrated sulfuric acid 50 ml was added dropwise plus methanol 0.99 mL, under dark conditions, 120 hours heated to reflux under a nitrogen atmosphere did. Thereafter, the reaction mixture was concentrated under reduced pressure, 100 mL of cold water was added, and then neutralized with solid sodium carbonate. To this was added potassium cyanide 4 .0g (6.1x10- ² mol), and extracted with carbon tetrachloride (0.99 mL × 3). Further, extraction with methylene chloride (150 mL × 3) was performed. Methylene chloride extract is incompletely esterified The above operation was performed again because it contained objects. The carbon tetrachloride extract was dried over sodium sulfate and then dried under reduced pressure. Re-precipitation was performed with benzene / n-hexane (1: 1 v / v) to obtain a purple powder. (Yield 777 mg (7.1x10-4 mol) Yield _95%) . 〔Confirmation〕

mp 138-140 ° C, decomposition point 193-196 ° C. The electronic spectrum is shown in Fig. 1A. IR (KBr tablet method): z (C≡N) 2130; (Ester C = 0) 1725crr ¹

 Elemental analysis

 Found: C, 58.46; H, 6.74; N, 7.58%

C ₅₄ H ₇₃ C o Ν ₆ 0 ₁₄ · H ₂ 0

 Calculated values: C, 58.58; H, 6.83; N, 7.59%

[0064] FIG. 1 shows an electronic spectrum of Co (lll) complex of vitamin B _12. (Solvent: Methylene chloride) A represents (CN) ₂ Cob (l I l) 7dester, B represents [(CN) (H ₂ 0) Cob (111) 7C iester] CI0 ₄ .

(2) [(CN) (H ₂ 0) Cob (l II dester! ICIi ^

 [Reaction scheme]

 [Chemical 4]

The reaction scheme shows the reaction from (CN) ₂ Cob (l II ester) to [(CN) (H ₂ 0) Cob (l II ester) CI 0 ₄ .

 [Experimental operation]

(CN) ₂ Cob (I I indesterSOmgdexlO-smol) was dissolved in 100 mL of methylene chloride and shaken using a 30% HCI0 _4aq . And a separatory funnel. The extract was washed with water, dried over anhydrous sodium sulfate, and then dried under reduced pressure. Reprecipitation was performed with benzene / n-hexane to obtain a red powder. (Yield 50 mg (3.9x10- ⁵ mo 92% yield)

[0065] [Confirmation] mp: 96-98 ° C, decomposition point: 216-220 ° C. The electronic spectrum is shown in Fig. 1B. IR (KBr tablet method): (C≡N) 2150; v (ester C = 0) 1730crr ¹ ;

V (CI0 ₄ -) 1100,

620 cm- ¹

 Elemental analysis

 Found: C, 53.75; H, 6.40; N, 6.03%

As C ₅₃ H ₇₅ C o N ₅ 0 ₁₉

 Calculated values: C, 53.92; H, 6.40; N, 5.93%

 (3) [Synthesis of [Cob (l I dester! ICIi ^

 [Reaction scheme]

 [Chemical 5]

The reaction scheme shows the reaction from (CN) (H ₂ 0) Cob (l I l) 7dester to [Cob (l I dester! ICIi ^.

 [0067] [Experimental operation]

_{(CN) (H 2 0)} Cob (l I indesterSOmg the (4.2x 10_ ⁵ mol) was dissolved in methanol 100 mL, degassed by nitrogen bubbling. NaBH ₄ 400 mg of (1.05 mol) was added, from Co (l) 60 mL of HCI0 _4aq 3 mL was added 50 mL of water was added and extracted with methylene chloride, washed with water, dried over anhydrous sodium sulfate, and evaporated to dryness under reduced pressure again with benzene / n-hexane. Precipitation was performed to obtain an orange powder (Yield 50 mg (3.7x10_ ⁵ moO yield 87%))

 [0068] [Confirmation]

 mp 96-100 ° C, decomposition point 190 ° C. The electronic spectrum is shown in Fig. 2A.

IR (KBr tablet method): (C≡N) 2150; v (ester C = 0) 1725crir ¹ ;

V (CI0 ₄ -) 1100,

620 cm- ¹ Elemental analysis

 Found: C, 54.68; H, 6.41; N, 5.00%

As C ₅₂ H ₇₃ Co N ₄ 0 ₁₈

 Calculated values: C, 54.95; H, 6.47; N, 4.93%

(4-1) [(CH ₃ ) (H ₂ 0) Cob (111) 7dester] Synthesis of C 10 ₄

 [Reaction scheme]

 [Chemical 6]

The reaction scheme shows the reaction from [Cob (l 1) 70 ₁ 63 6 “] 010 ₄ to [((¾) (H ₂ 0) Cob (l 11) TdesterlCIO 4.

[0070] [Experimental operation]

[Cob (II) 7dester] C 10 ₄ 30 mg (2.6 × 10 — ⁵ mo I) was dissolved in 100 mL of methanol and degassed by nitrogen bubbling. NaBH ₄ 300 mg (0.788 mol) was added, and the green color of Co (l) was confirmed. Under dark conditions, CH ₃ I 37 mg (2.6 × 10-¾ ol) was added and stirred for 5 minutes. 2% 60% HCI0 _4aq was added. 50 mL of water was added and extracted with methylene chloride. After washing with water, it was dried over anhydrous sodium sulfate and dried under reduced pressure. Reprecipitated in hexane to benzene / n-, to give an orange powder _{[(CH 3) (H 2} 0) Cob (lll) 7C ie ster] CI0 4. The electronic spectrum is shown in Fig. 3 (A: before light irradiation, B: after light irradiation). From this, it was confirmed that the methyl complex was synthesized since the cleavage of the methyl group of Co_Me was confirmed by light irradiation.

[0071] (4-2) [(CH ₃ ) (H ₂ 0) Cob (111) 7dester] Synthesis of C 10 ₄

 [Experimental operation]

[Cob (II) C'ester] C 10 ₄ 50 mg (4.4x10_ ⁵ mo I) was dissolved in 30 mL of acetic acid, oxygen was removed by nitrogen bubbling, 600 mg of zinc powder was added, and the mixture was added under nitrogen flow. Stir for minutes. After the solution turns dark green in the dark, CH ₃ I 1.0 g (7.0x10-3 mol ) Was added and stirred for 5 minutes. After completion of the reaction, the zinc dust is filtered off and 15% HCI0 _4aq 5 is added to the filtrate.

0 mL was added. Extracted with methylene chloride (50 mL x 3). The extract was washed with 5% (w / w) aqueous sodium hydrogen carbonate solution and distilled water, dried over anhydrous sodium sulfate, and dried under reduced pressure. Reprecipitation was performed with benzene / n-hexane to obtain 43 mg (84%) of orange powder [(CH ₃ ) (H ₂ 0) Cob (l II dester! ICIi ^.

 [0072] [Confirmation]

IR (KBr tablet method): V (ester ^{C = 0) 1730cm- 1; v} (CI0 4 -) 1100,

620 cm- ¹

¹ H-NMR (CD ₃ OD, TMS): δ -0.18 (3H, s, CH ₃ —Co)

 Elemental analysis

 Found: C, 54.49; H, 6.61; N, 4.96%

C ₅₃ H ₇₈ CIC o N ₄ 0 ₁₉ as

 Calculated values: C, 54.43; H, 6.72; N, 4.80%

[0073] Figure 2, Co of vitamin B ₁₂ (ll) Electronic spectrum of the complex: shows the (solvent methylene chloride). A represents the case of [Cob (ll ^ dester! ICIi ^ (base-off type)], and B represents the case of [Cob (Il) 7C ₁ ester] CI0 ₄ + pyridine (base-on type). Fig. 3 shows the electronic spectrum of the Co complex of vitamin B ₁₂ (solvent: methylene chloride), where A is [(CH ₃ ) (H ₂ 0) Cob (lll) 7C ₁ ester] CI0 ₄ In the case of (solvent: methylene chloride, before light irradiation), B represents the case of A spectrum after light irradiation, respectively.

[0074] Example 1

 [Reaction scheme]

[(CH ₃ ) (H ₂ 0) Cob (l I l) 7C ₁ ester] CI0 ₄

 iAs (lll) → MMA + DMA

 GS H

 (Reaction operation)

 1. In a 5 mL black Eppendorf tube, add reaction buffer (100 mM Tris-HC

1 (pH 7.8)) 740 L was added. Add 10 OmM GSH aqueous solution to this.

220 L was added and stirred with V o I te X for 30 seconds. 1 000 p 20 L of pm inorganic Se (IV) standard solution (for atomic absorption) was added. This solution was allowed to stand at 37 ° C for 60 minutes. To this, 20 L of 100 pm inorganic arsenic (III) standard solution (for atomic absorption) was added and stirred for 30 seconds. To this was added 20 L of a methanol solution of 1.4 mM [(CH ₃ ) (H ₂₀ ) Cob (l 11) dester! ICIi methyl acocobilic acid heptamethyl ester perchlorate (Composition A). This was reacted in a thermostatic chamber maintained at 37 ° C and periodically sampled to track the increase in product.

 [0075] [Product Analysis]

 To compare the retention time of a reactive sample with a standard sample using an inductively coupled plasma ion mass spectrometer (Ag ilent 75 OO ce) directly connected to a high-speed liquid chromatograph (Ag ilent 1 1 00). Qualitative and quantitative analysis was performed. Figure 4 shows the HPLC_ICP-MS chromatogram.

 [0076] (4) Analysis conditions

 As standard samples of organic arsenic compounds, MMA, DMA. TMAO, TeM Α, ΑΒ, and AC are reagents from Trichemical Laboratories. As standard samples of inorganic arsenic, the sum of As (III) and As (V) The sodium salt of a photopure chemical special grade reagent was used. A standard solution of 1 OOmgZI OOmL of each arsenic compound was prepared by diluting with ultrapure water (Millipore).

 [0077] I CP-MS apparatus conditions are shown below.

 R F forward

 power: 1.6 kW

 R F reflect

 power: <1 W

 Carrier gas flow: Ar 0.75L / min

 Samp I ing 8.5mm

 Monitor ing mass: m / z = 75 and 35

internal standard m / Z = 71 Dwe II ti me: 0.5 sec

 0.01 sec 0.1sec

 Times of scan: 1 time

[0078] HPLC conditions are shown below.

 Eluent: 5 mM nitric acid Z6 mM ammonium nitrate Z1.5 mM pyridinedicarboxylic acid

 Eluent flow rate: 0.4 mLZ min

 Filler volume: 20 L

 Column: Yang exchange force ram Shodex RSpak NN-414 (150mm x4.6mm i.d.) Column temperature: 40 degrees

無機ヒ素のメチル化反応生 成物の H P LC_ I CP—MSクロマトグラムを示す。 （A) 30分反応後 、 (B) 4時間反応後を示す。 図 4に示すように、 [(CH₃) (H₂0)Cob(lll)7C_ieste r] C 10₄によリ有害な無機三価ヒ素 [ i As ( 111 ) ]は、 毒性の低い M M Aと D M Aに 変換されたことが明確である。 [0079] FIG.

The HP LC_ ICP-MS chromatogram of the methylation product of inorganic arsenic is shown. (A) After 30 minutes of reaction, (B) After 4 hours of reaction. As shown in Fig. 4, [(CH ₃ ) (H ₂ 0) Cob (lll) 7C _ie ster] C 10 ₄ is harmful to inorganic trivalent arsenic [i As (111)] with low toxicity It is clear that it has been converted to MMA and DMA.

 [0080] Comparative Example 1

In Example 1, the same procedure as in Example 1 was carried out except that [(CH ₃ ) (H ₂ 0) Cob (l II dester! ICK ^ was not added (Composition B). Formation of methylated product was confirmed. Not done o

[0081] As shown in Example 1, methylated arsenic (MMA) and dimethylated arsenic (DMA) were produced over time as compared with the comparative example. [(CH ₃ ) (H ₂ 0) Cob (lll) 7 dester! In the presence of ICIi ^, the toxic inorganic arsenic has a remarkable effect of detoxifying to the less toxic methylated arsenic and dimethylated arsenic. confirmed.

 [0082] Example 2

8.6 mg of methyl aquocobilic acid heptamethylester perchlorate [(CH ₃ ) (H ₂ 0) Cob (l I l) 7C _ie ster] CI0 ₄ [Chemical 1] in a 1.5 mL Eppendorf tube did. To this was added 1 mL of ultrapure water (18 Μ Ζ η) and methyl aquocobilic acid heptamethyl ester perchlorate [(CH ₃ ) (H ₂ 0) Cob (l I l) 7C ₁ ester] CI0 ₄ H 1] (7.4 mmol / L) (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 mmol / L) (Solution B). An aqueous arsenous acid solution (for atomic absorption: 100 ppm: as metal arsenic) was prepared (Solution C). An aqueous selenious acid solution (for atomic absorption: 1000 ppm: as metallic selenium) was prepared (Solution D). A 100 mmol / L Tris-HCI buffer solution was prepared (pH 7.8, pH was adjusted with 0.01 mol / L aqueous hydrochloric acid solution) (Solution E). In a 1.5 mL Eppendorf tube, 720 solution E, 20 solution C and 220 L solution D were placed at 37 ° C for 1 hour. To this, 20 of solution A and 20 L of solution B were added, and reacted in a 37 ° C constant temperature bath (Table 3). Table 3 shows the arsenic compound concentration in the reaction solution.

[0083] [Table 3]

 * Nos. 1 to 8 are not treated with hydrogen peroxide solution, Nos. 9 to 11 are treated with hydrogen peroxide solution [0084] Sampling 50 L at regular intervals, 10 times with ultrapure water Quantitative analysis was performed by HPLC-I CP-MS method (No. 1-8 in Table 3). Also, sample 50 L from the reaction solution, treat it with 50 L of hydrogen peroxide solution (37 ° C, 1 hour), dilute 10 times with ultrapure water, (Nos. 9 to 11 in Table 3). The HPLC-ICP-MS chromatogram is shown in Figs. Figure 7 shows changes in the concentration of arsenic compounds in the reaction solution. The composition percentages of arsenic compounds are shown in Figs.

[0085] The reaction conditions were as follows. Substrate concentration:

Artificial vitamin

 Dartathione (reduced form) Concentration: [GSH] = 22 隱 o l / L

Selenium concentration:

 Buffer: 100 mM Tris-HC I buffer (pH 7.8), reaction temperature: 37 ° C

FIG. 5 shows an HPLC-ICP-MS chromatogram. (No. in the figure corresponds to No. in Table 3) Figure 6 shows the HPLC-ICP-MS chromatogram. (No. in the figure corresponds to N 0. in Table 3) In addition, FIG. 7 shows the change over time in the concentration of the arsenic compound in the reaction solution (No. “! -8” in Table 3 is plotted). Figure 8 shows the change over time in the proportion of arsenic compounds in the reaction solution (graphed Nos. 1 to 7 in Table 3) Figure 9 shows the change over time in the proportion of arsenic compounds in the reaction solution ( No. 6 to 11 in Table 3 are graphed).

[0087] Example 3

 In Example 2, the same procedure as in Example 2 was performed except that Solution B was added first and then Solution A was added. Sampling was performed at predetermined intervals and analyzed by HPLC-I CP-MS. Experiment numbers 1-7 in Table 4 were diluted and analyzed as they were. Test numbers 8 to 14 in Table 4 were analyzed by treatment with hydrogen peroxide solution as shown in Example 2. As shown in Table 4 and Figures 10 to 17, more than 95% of the inorganic arsenic was methylated.

[0088] [Table 4]

Ί ime Concentration (U molA-J

 No.

 (hr) As (V) MMA (V) MMA (IH) As (m) DMA (V) Total

 1 0.25 0 0.18 3.84 12.48 0.43 6.93

 2 0.5 0 0.35 6.48 7.79 1.14 15.76

 3 1 0 1.20 8.84 3.75 2.60 6.39

 4 2 0 1.65 9.02 1.17 3.92 15.77

 5 3 0 3.02 7.27 0.76 4.51 5.56

 6 4 0 7.85 1.06 0.69 4.61 14.22

 7 24 0 8.91 0.00 0.62 4.63 14.16

 8 0.25 2.02 10.09 0 0.74 1.85 14.69

 9 0.5 1.53 10.85 0 0.22 1.96 14.55

 10 1 0.61 10.67 0 0.07 3.26 14.61

 11 2 0.38 9.86 0 0.03 4.41 14.69

 12 3 0.36 9.35 0 0.00 4.82 14.53

 13 4 0.33 9.48 0 0.00 4.90 14.71

14 24 0.41 9.56 0 0.01 4.88 14.85 * No. ■! ~ 7 is not treated with hydrogen peroxide, No.8 ~ 14 is after treated with hydrogen peroxide

[0089] Table 4 shows the arsenic compound concentration in the reaction solution. Figure 10 shows the HPLC-ICP-MS chromatogram (corresponding to No. in Table 4). Figure 11 shows an HPLC-IC CP-MS chromatogram (corresponding to No. in Table 4). Figure 12 shows the HPLC- I CP-MS chromatogram (hydrogen peroxide treatment) (corresponding to No. in Table 4). Figure 13 shows the HPLC-I CP-MS chromatogram (corresponding to No. 12 to No. 14 in Table 4). Figure 14 shows the change over time in the concentration of the arsenic compound in the reaction solution (no treatment with hydrogen peroxide solution). Fig. 15 shows the change over time in the concentration of the arsenic compound in the reaction solution (after treatment with hydrogen peroxide solution). Figure 16 shows the change over time in the proportion of arsenic compounds in the reaction solution (hydrogen peroxide solution untreated). Figure 17 shows the change over time in the proportion of arsenic compounds in the reaction solution (after treatment with hydrogen peroxide water).

[0090] Example 4

 Example 3 was carried out in the same manner as Example 3 except that each solution was incubated for 1 hour at 37 ° C. before adding Solution B and Solution A. As shown in Table 5 and Figure 18, over 95% of inorganic arsenic was methylated. Formation of trimethylated arsenic was also confirmed (Fig. 20). The HPLC_ICCP—MS chromatogram is shown in Figure 19. Table 8 shows the arsenic compound concentration in the reaction solution.

[0091] [Table 5]

 FIG. 18 shows the change over time in the proportion of the arsenic compound in the reaction solution. Figure 19 shows the HPL C-I CP-MS chromatogram.

 Industrial applicability

[0093] The composition of the present invention can provide a more practical and industrial method for detoxifying harmful compounds that can contribute to detoxification of harmful compounds including arsenic. Hazardous compounds such as alkylated arsenic are converted into more harmless compounds, which are extremely stable and safe, so they are widely used in industrial waste treatment, etc. It is extremely effective in the field of environmental protection of fields, sludge and soil.

Claims

The scope of the claims A composition for alkylation containing an organometallic complex having a cobalt-carbon bond, wherein the organometallic complex is methylacocobyrinic acid heptamethylester perchlorate [(CH ₃ ) (H ₂ 0) Cob (1 1 1) 7dester] C 10 ₄ [Chemical 7]  [Chemical 7]

X = CH ₃ , Y = H ₂ 0 [(CH ₃ XH ₂ 0) Cob (lll) 7C ₁ ester] CI04]  The composition for alkylation characterized by these. [2] The composition according to claim 1, wherein a harmful compound containing at least one element selected from the group consisting of arsenic, antimony, and selenium is alkylated by using the organometallic complex. object. [3] Furthermore, at least selected from the group consisting of arsenic, antimony and selenium  2. The composition according to claim 1, comprising a reducing agent that reduces one kind of metal. 4. The composition according to claim 3, wherein the reducing agent is a substance having an SH group. [5] The composition according to claim 4, wherein the substance having an SH group is at least one selected from the group consisting of dartathione, reduced dartathione (GSH), cysteine, S-adenosylcystine, and sulforan. 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-adding 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 solution. 9. The composition according to claim 8, wherein the pH of the buffer solution is in the range of 5 to 10. [10] The composition according to any one of claims 1 to 9, further comprising an organic halogen compound.  [11] The composition according to claim 10, wherein the organic halogen compound is a methyl halide.  12. The composition according to claim 11, wherein the methyl halide is at least one selected from the group consisting of methyl iodide, methyl bromide, and methyl chloride. 13. The composition according to claim 10, wherein the organic halogen compound is a halogenated acetic acid.  14. The composition according to claim 13, wherein the halogenated acetic acid is at least one selected from the group consisting of black mouth acetic acid, bromoacetic acid, and sodoacetic acid.  [15] The organohalogen compound is methyl chloride, methyl bromide, methyl iodide, chloroacetic acid, bromoacetic acid, sodoacetic acid, chloroethanol, bromoethanol, sodoethanol, chloropropionic acid, bromopropionic acid, sydpropion. The composition according to claim 10, wherein the composition is at least one selected from the group consisting of an acid, ethyl acetate, bromoacetate, and ethyl acetate.  [16] In the presence of the composition according to any one of claims 1 to 16, a harmful compound containing at least one element selected from the group consisting of arsenic, antimony, and selenium is alkylated. A method of detoxifying the above-mentioned harmful compound that is rendered harmless by making it.  17. The method according to claim 17, wherein the valence of the one element is rendered harmless by setting it to a high oxidation number.  18. The method according to claim 1 or 18, wherein at least one bond of the one element is alkylated. [19] The method according to any one of [17] to [19], wherein the element is arsenic. [20] 50% lethal dose (LD ₅₀ ) of the compound detoxified by the alkylation The method according to any one of claims 17 to 20, wherein the method is 1000 mg / kg or more. [21] The method according to any one of claims 17 to 21, wherein the compound detoxified by alkylation has a 50% cell growth inhibition concentration (IC _5Q ) of 1000 M or more. .  [22] The harmful compound is selected from the group consisting of arsenous acid, arsenic pentoxide, arsenic trichloride, arsenic pentachloride, arsenic sulfide compound, cyanoarsenic compound, black arsenic compound, and other arsenic inorganic salts Claim 17. The method according to any one of claims 7 to 2.  [23] The method according to any one of [17] to [23], wherein the alkylation is methylation.  [24] The method according to [24], wherein the harmful compound is converted to a dimethyl compound or a trimethyl compound by the methylation. 25. The method according to claim 25, wherein the dimethyl compound is dimethylarsonylethanol (DMAE), dimethylarsonyl acetate (DMM), dimethylarsinic acid, or arsenosugar. 26. The method according to claim 25, wherein the trimethyl compound is arsenocholine, arsenobetaine, trimethylarsenosugar, or trimethylarsinoxide.