WO2007011251A1

WO2007011251A1 - Method for neutralising a toxic agent into safe non-toxic products

Info

Publication number: WO2007011251A1
Application number: PCT/RU2005/000384
Authority: WO
Inventors: Viktor Stanislavovich Polyakov; Valery Vasilievich Ermilov; Sergey Ivanovich Malekin
Original assignee: Zakrytoe Aktzionernoe Obschestvo 'elit Holding'
Priority date: 2005-07-20
Filing date: 2005-07-20
Publication date: 2007-01-25

Abstract

The invention relates to toxic agent processing engineering. The inventive method for neutralising toxic agents consists in interacting the toxic agents with a detoxicating reagent, in neutralising the thus obtained product in such a way that the pH thereof ranges from 7 to 8, thereby ensuring the subsequent product biodegradation, in carrying out the biodegradation in such a way that a harmless biomass and waste waters are obtained. The detoxicating reagent is embodied in the form of a composition containing at least one type of natural α amino acid, wherein the ratio between the toxic agent and the detoxicating reagent is equal to 1:1-1:3 and the interreaction is carried out by mixing at a temperature of 20-45 °C for a time of 0.5-1 hour. Said invention makes it possible practically totally neutralise the toxic agent under soft conditions with the aid of a low-cost and accessible reagent.

Description

A method of neutralizing toxic substances into safe non-toxic products.

FIELD OF THE INVENTION The invention relates to a technology for processing toxic substances (OB).

In January 1993, representatives of more than 130 states signed the final act of the Chemical Weapons Agreement, which prohibits the production, use, sale and storage of all types of chemical weapons and their means of delivery, and envisages the destruction of existing stockpiles of chemical weapons by 2005. In Russia, in accordance with international obligations, by April 29, 2003, 1% of Russian chemical weapons should be destroyed, which amounts to 400 tons of poisonous substance.

Prior art Throughout the world until the seventies of the twentieth century, chemical weapons were destroyed by methods that are now prohibited: burning in the open air, evaporation and dilution in airspace, flooding in the seas and oceans, burial in the ground. In the seventies, they began to use the neutralization of toxic substances (OB) using chemicals and the subsequent burning of neutralization products.

However, due to the difficulties encountered in the implementation of these processes, the management of the chemical weapons destruction program in the United States in the 90s focused on direct burning of OB in specially equipped plants. At the same time, public protests and aggravated technical problems at OB burning facilities led to the need to seek

SUBSTITUTE SHEET (RULE 26) new technologies for the disposal of OB.

The prior art method for processing pollutants; the environment (US 6,039,882). The method and composition are intended for the processing of substances that pollute the environment and are in the soil, in sediments, in aquifers, water. In addition, the method and composition described in the patent can be used to treat containers containing contaminants. The essence of the method is the reaction of a pollutant with a composition that includes a metal and a compound containing sulfur, resulting in environmentally acceptable chemically less active products. The method is applicable for contaminants such as halogenated hydrocarbons, for pesticides, for chemical weapons and for dyes. The composition in a preferred embodiment contains powdered iron and iron sulfide. The addition of alcohol to the composition increases the efficiency of the processing reaction, especially for soil contaminants and sediments. There is also known a method of processing a toxic substance by hydrolysis (US

5678243). The method includes adding water to the chemical reagent (OB) so that the hydrolysis reaction of the chemical reagent with water occurs in the molar ratios indicated in the patent. In a preferred embodiment, the method is carried out in the field in containers in which the processed substance (aminoalkylphosphonothiolates) is stored. The method includes mixing the contents of the container after adding water.

There is also a method of processing chemical weapons by alkaline hydrolysis (WO 9718858). The method includes reacting a chemical weapon with a nitrogen-containing base, such as anhydrous liquid ammonia, in which an active metal, for example, sodium, is dissolved.

SUBSTITUTE SHEET (RULE 26) The disadvantage of the methods described in the above sources of information is their inconsistency with modern requirements for environmental safety of the destruction of chemical weapons (toxic substances) and pesticides.

The US National Research Committee recommended for further development and implementation of an alternative technology for neutralizing OBs with alkaline hydrolysis, followed by biodegradation of the resulting less toxic hydrolysis products.

Biodegradation of chemicals is a natural process that improves the environment. Scientists are also trying to obtain microorganisms and enzyme systems for direct biodegradation of OB. However, due to the high toxicity of OBs, microbial cells are still capable of destroying OBs only at the cost of their lives, which significantly increases the cost of biodegradation of OBs and makes them highly controversial from an environmental point of view.

Separately isolated enzyme systems are too specific (that is, each type of toxic substance needs its own enzyme), are whimsical and expensive for practical use in the direct destruction of OB.

The closest analogue to the invention is a method for processing a toxic substance (mustard gas) by alkaline hydrolysis followed by biodegradation of less toxic substances formed as a result of hydrolysis (US 6017750). The method involves the processing of a mixture of 2,2'-dichlorodiethylcylphide and bis-2-2-chloroethylthioethyl ether (HT), the method comprising the following operations: combining HT with a hydrolyzing agent to form a mixture of thiodiglycol (TDG) and bis-2- ( 2-hydroxyethylthio) ethyl ester (s) (T-OH); - neutralizing the aforementioned mixture of TDG and T-OH to a pH that provides

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SUBSTITUTE SHEET (RULE 26) the possibility of further biodegradation of the mixture; biodegradation of the aforementioned neutralized TDG and T-OH mixtures to obtain neutralized biomass and wastewater.

The disadvantages of the closest analogue include obtaining at the 1st stage (alkaline hydrolysis) toxic toxic products of neutralization, which significantly complicates and increases the cost of all processes associated with their further storage, transportation, etc., as well as increases the cost of their subsequent biodegradation, since special installations for biodegradation and continuous renewal of microorganisms that die due to the high toxicity of alkaline hydrolysis products are required.

Disclosure of the invention

The technical result of the invention is the completeness of detoxification, during which the toxicity of the initial OB is reduced so that it becomes possible further biodegradation of detoxification products. The technical result is achieved by the method of neutralizing poisonous substances and Vx hydrolyzate set forth in the claims, based on their interaction with an amino acid reagent (AKP). Amino acid reagent must contain at least one of the natural α-amino acids, such as alanine, arginine, aspartic acid, asparagine, valine, histidine, glycine, glutamic acid, glutamine, isoleucine, leucine, lysine, methionine, proline, series, tyrosine, threonine , tryptophan, phenylalanine, cysteine (see table 1). Amino acid reagent can also be obtained by alkaline hydrolysis of protein-containing industrial waste.

The low-toxic products resulting from the interaction of an amino acid reagent with an OB or Vx hydrolyzate are neutralized to a pH of 7-8, which enables their further biodegradation. Biodegradation is carried out

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SUBSTITUTE SHEET (RULE 26) in the traditional way using microorganisms, including from "activated sludge", until neutralized biomass is obtained.

The described method does not have the disadvantages of the above methods of detoxification of OB and has in comparison with them a number of the following advantages: - ensuring the destruction of OB or hydrolyzate Vx with subsequent bio-utilization of their decomposition products, that is, with the possibility of

product obtained as a result of processing by microorganisms, directly as soil improving additives for lands of wide purpose up to

agricultural;

- the possibility of applying the developed method for land rehabilitation and

objects infected with OB;

- the use of a highly effective and affordable reagent for detoxification

OB;

- significantly lower toxicity of neutralization products compared with known technologies that use alkaline hydrolysis, products

neutralization of OB amino acid reagent can be disposed of

microorganisms in the process of their life, therefore, significantly

the range of microorganisms capable of processing neutralized

the proposed method OB; - the ability to transport neutralization products formed on

the first stage, as low-toxic waste, and the possibility of their biodegradation in ordinary enterprises of biochemical wastewater treatment.

Examples of carrying out the invention

Example 1. Detoxification of mustard gas under the action of an amino acid reagent.

A mustard sample (2,2'-dichlorodiethyl sulfide) was prepared according to the method

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SUBSTITUTE SHEET (RULE 26) Meyer from 2,2'-dioxiethylcylphide and concentrated hydrochloric acid, isolated by double distillation. Indicators: boiling point 95-97710 mm RT. Art., d ²⁰ _n = 1.275, n ² ° _D ⁼ 1.528 correspond to published data.

Mustard detoxification was carried out according to the following method. In a 50 ml Erlenmeyer flask equipped with a magnetic stirrer, a solution of 80 mg of mustard gas in 10 ml of 0.1 M amino acid reagent was placed, which corresponds to their ratio of 1

: 2, and kept under stirring at a temperature of 4O ⁰ C for 1 hour. The solution was adjusted to pH = 7.0 and transferred to the study of acute toxicity after intragastric administration in experiments on rats; the dose of LD ₅₀ and detoxification products exceeded 5000 mg / kg.

Morphological studies of internal organs, conducted 14 days after the introduction of detoxification products, showed a complete absence of any manifestations of toxic effects. Mutagenic and carcinogenic properties were not found in detoxification products (Ames test with metabolic activation - Salmonella / microsome, strains TA-98 and TA-100).

Example 2. Detoxification and biodegradation of lewisite under the action of an amino acid reagent and microorganisms.

A sample of a-lewisite (2-chlorophenyl dichloropapcine) was prepared by the interaction of arsenic trichloride with acetylene in the presence of aluminum chloride and was isolated by double distillation. Indicators: boiling point 75-79 / 12 mm RT. Art., d ²⁰ n = 1,875, n ²⁰ _D = 1,06092 correspond to literature data.

The detoxification of lewisite was carried out according to the following method. In a 50 ml Erlenmeyer flask equipped with a magnetic stirrer, a solution of 124 mg of lewisite and 12 ml of 0.1 M amino acid reagent (pH 7 - 12) was placed, the ratio of reagents was 1: 2, kept under stirring at room temperature for 0.5 hour,

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SUBSTITUTE SHEET (RULE 26) pH adjusted to 7.0 and the solution was passed to evaluate acute toxicity in case of intra-gastric administration in experiments on rats rer OR lethal dose LD ₅₀ of the detoxification products exceeded 5000 mg / kg.

The same solution (reaction mass after detoxification of lewisite with 0.1 M solution of amino acid reagent) was studied as a substrate for the activity of bacteria strottosossus thermorhulis and activated sludge. Both the initial solution and diluted to concentrations of 10 ^" ÷ 10 ^{" 3} were used. 10 ml of solution was placed in test tubes and 2 ml of bacterial cultures with a titer of 10 ⁶ were added, then the tubes were incubated for one day at 37 ° С. day marked bacterial growth in all test tubes. Samples with the initial concentration of the reaction mass were transferred to the study of acute toxicity during intra-gastric administration in experiments on rats. It was found that the regal dose of LD50, solutions with stractosossus thermorhulis and activated sludge strains exceeds 5000 mg / kg. When studying all dose levels, no clinical manifestations of intoxication were detected. Morphological studies of the internal organs of animals, carried out 14 days after the introduction of detoxification products, showed a complete absence of any manifestations of toxic effects. Mutagenic and carcinogenic properties were not found in detoxification products (Ames test with metabolic activation - Salmonella / microsome, strains TA-98 and TA-I 00). Example 3. Detoxification of lewisite under the action of lysine.

In a 30 ml Erlenmeyer flask equipped with a magnetic stirrer, 62 mg of a-lewisite identical to the lewisite used in Example 1 and 15 ml were placed

0.1 M sodium lysinate solution (pH 8 - 12), reagent ratio 1: 3. The mixture was kept under stirring at room temperature for 0.5 hours, the pH of the solution was adjusted to 7.0 and transferred to the study of acute toxicity, at

SUBSTITUTE SHEET (RULE 26) intragastric administration in experiments on rats, a re-lethal dose of LD ₅₀ , of detoxification products exceeds 5000 mg / cr. Morphological studies of internal organs, conducted 14 days after the introduction of detoxification products, showed a complete absence of any manifestations of toxic effects. Mutagenic and carcinogenic properties were not found in detoxification products (Ames test with metabolic activation - Salmonella / microsome, strains TA-98 and TA-100). Example 4. Detoxification of lewisite under the action of cysteine.

In a 30 ml Erlenmeyer flask equipped with a magnetic stirrer were placed 62 mg of a-lewisite identical to the lewisite used in Example 2, and 12 ml of 0, IM cysteine sodium salt solution (pH 8-12), which corresponds to the ratio

13. The mixture was kept under stirring at room temperature for 0.5 hours, the pH of the solution was adjusted to 7.0 and transferred to an acute gastric toxicity study in experiments on rats, a re-lethal dose of LDs ₀ , and detoxification products in excess of 5000 mg / kg. Morphological studies of internal organs, conducted 14 days after the introduction of detoxification products, showed a complete absence of any manifestations of toxic effects. Mutagenic and carcinogenic properties were not found in detoxification products (Ames test with metabolic activation - Salmonella / microsomes, strains TA -98 and TA-100). Example 5. Detoxification of sarin by the action of an amino acid reagent.

Detoxification of sarin was carried out according to the following method. In a 50 ml Erlenmeyer flask equipped with a magnetic stirrer, a solution of 100 mg of sarin in 14.3 ml of 0.1 M amino acid reagent (1: 2 ratio) was placed under stirring at room temperature for 0.5 hours. The solution was adjusted to pH = 7.0 and transferred to the study of acute toxicity with intragastric

SUBSTITUTE SHEET (RULE 26) In the experiments on rats, the lethal dose of LDso, the detoxification products exceeded 5000 mg / kg.

Morphological studies of internal organs, conducted 14 days after the introduction of detoxification products, showed a complete absence of any manifestations of toxic effects. Mutagenic and carcinogenic properties were not found in detoxification products (Ames test with metabolic activation - salmonella / microsomes, strains TA -98 and TA-100).

Example 6. Detoxification of soman by the action of an amino acid reagent.

Zoman detoxification was carried out according to the following procedure. In a 50 ml Erlenmeyer flask equipped with a magnetic stirrer, a solution of 100 mg of soman in 10.9 ml of 0.1 M amino acid reagent (1: 2 ratio) was placed under stirring at room temperature for 0.5 hours. The solution was adjusted to pH = 7.0 and transferred to the study of acute toxicity after intragastric administration in experiments on rats, a re-lethal dose of LD ₅₀ , and detoxification products exceed 5000 mr / kg.

Example 7. Detoxification of soman under the action of phenylalanine.

Zoman detoxification was carried out according to the following procedure. Into the flask

50 ml Erlenmeyer, equipped with a magnetic stirrer, placed the solution

450 mg of soman in 25 ml of a 0.1 M solution of phenylalanine sodium salt (1: 1 ratio) was kept under stirring at room temperature for 0.5 hours.

SUBSTITUTE SHEET (RULE 26) The solution was adjusted to pH = 7.0 and transferred to the study of acute toxicity after intragastric administration in experiments on rats. LD ₅ 0, by mouth, detoxification products exceed 5,000 mg / kg.

Example 8. Detoxification of soman by sodium glycinate. Zoman detoxification was carried out according to the following procedure. Into the flask

A Erlenmeyer with a capacity of 50 ml equipped with a magnetic stirrer was placed a solution of 450 mg of soman in 50 ml of 0.1 M sodium glycinate solution (1: 2 ratio) was kept under stirring at room temperature for 0.5 hours. The solution was adjusted to pH = 7.0 and transferred to the study of acute toxicity after intragastric administration in experiments on rats, a re-lethal dose of LD ₅₀ , detoxification products exceed 5000 mg / kg.

Morphological studies of internal organs, conducted 14 days after the introduction of detoxification products, showed a complete absence of any manifestations of toxic effects. Mutagenic and carcinogenic properties were not found in detoxification products (Ames test with metabolic activation -

Salmonella / microsome, strains T A-98 and T A-100). Example 9. Detoxification and biodegradation of the hydrolyzate of substance Vx under the action of an amino acid reagent and microorganisms.

A sample of the hydrolyzate of substance Vx was obtained by hydrolysis of 0-

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SUBSTITUTE SHEET (RULE 26) isobutyl-8-2-diethylaminoethyl-methylthiophosphonate in water at a ratio of 7: 100 (by volume) for 3 months at room temperature. A study of acute toxicity during intragastric administration in experiments on rats showed that the lethal dose of LDs ₀ , hydrolyzate of substance Vx, is 750 mg / kg. The clinical picture of intoxication is curariform in depolarizing type.

The detoxification of the hydrolyzate of substance Vx was carried out as follows. A solution of 5. ml of a hydrolyzate of substance Vx, 10 ml of a 0.1 M amino acid reagent (pH 8 - 12) ratio and 35 ml of water was placed in a 100 ml Erlenmeyer flask equipped with a magnetic stirrer, stirred at room temperature for 0.5 hour. A study of acute toxicity during intra-gastric administration in experiments on rats showed that the re-lethal dose of LD _{50 of the} resulting reaction mass exceeds 5000 mg / kg.

Hydrolyzate Vx (reg. Lethal dose LD _5O , 750 mg / kg) was studied as a possible substrate for the life of a number of microorganisms (strottosossus thermorhulis, basillus subtilis, escherisia coli, strossossus lactis). Tubes containing 10 ml of Vx hydrolyzate were seeded (2 ml of culture each) with all 4 strains.

After a day of incubation at 37 ° C, all crops died.

The reaction mass obtained after detoxification of the hydrolyzate of substance Vx with an amino acid reagent was studied as a substrate for the activity of the bacteria strottosossus thermorhulis and activated sludge. Both the initial solution and dilution up to 10 ^{"1 were used} . 2 ml of bacteria cultures with a titer of 10 ^b were added to the tubes containing 10 ml of solutions; the tubes were incubated for 24 hours at 37 ^° C. Bacteria grew in of all test samples, samples with the initial concentration of the reaction mass were transferred to the study of acute and chronic toxicity during intragastric administration in experiments on rats.

SUBSTITUTE SHEET (RULE 26) It was found that the lethal dose of LDso, samples of the reaction mixture with both strain strossossus thermorhulis strain and activated sludge was found to exceed 5000 mg / kg. The clinical picture of intoxication is absent.

In experiments to determine chronic toxicity, rats received 5000 mg / kg daily for 5 days. Thus, the re-lethal dose of LDs ₀ exceeds 25,000 mg / kg. Morphological studies of the internal organs of animals, conducted 14 days after the removal of detoxification products, showed a complete absence of any manifestations of toxic effects. Mutagenic and carcinogenic properties were not found in detoxification products (Ames test with metabolic activation - Salmonella / microsomes, strains T A-98 and T AI 00).

A control study of the toxicity of an amino acid reagent (pH = 7.0) in a chronic experiment showed that the lethal dose LD ₅₀ exceeds 25,000 mg / kg (5,000 mg / kg daily for 5 days).

The table N≥l shows the amino acid composition of the amino acid reagent (AKP) from the table shows that during alkaline hydrolysis of industrial white waste, 15 of the 20 natural α-amino acids are preserved, and this corresponds to the data presented in the monograph of ALinger Biochemistry. M. Mir: 1976. Table N ° 2 shows the data of toxicometric measurements of the reaction mass obtained after the interaction of the OB with the amino acid reagent. From the data of the table it is seen that for all the studied OBs (mustard gas, lewisite, sarin, soman), these data for intragastric administration to rats is greater than 5000.0 mg / kg. Table N ° 3 shows the toxic effects of the Vx hydrolyzate and reaction mass after its interaction with AKP. The table shows that the toxicity of the Vx hydrolyzate is ~ 750 mg / kg when administered intravenously to rats, a curariform clinical picture. Immediately after interaction

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SUBSTITUTE SHEET (RULE 26) Vx hydrolyzate with AKP LDso of the reaction mass (rat, intragastric, mg / kg) is more than 5000.0 mr / kg, the clinical picture is absent. Data on direct and mutagenic activity of the reaction masses after detoxification with their amino acid reagent are shown in table N ° 4. The data in the table show that, against the background of the control aminoanthracene mutagen, the reaction masses obtained as a result of interaction with AKP do not exhibit noticeable mutagenic activity. The toxic effects of the reaction masses given in Table N ° 5 after detoxification of the Vx hydrolyzate with AKP (1st stage), then bio-utilized by microorganisms, indicate that the toxicity of these reaction masses is higher than the 4th hazard class. Although the essence of the invention is described in detail in several examples of carrying out the invention, it should be understood that this is done only for illustration, and the invention is not limited to the listed examples of implementation.

The temperature conditions indicated in the claims and the reaction time are interconnected. Specific modes are selected based on the desired reaction rate and production capabilities.

The minimum reaction time is determined by the time of neutralization of the toxicity of the ecotoxicant. The increase in exposure time, regardless of the temperature at which the reaction is carried out, does not affect the result - neutralization of toxicity.

Thus, the invention allows to neutralize toxic substances - mustard gas, lewisite, sarin, soman and Vx hydrolyzate under mild conditions using a cheap and affordable reagent - a composition containing a mixture of natural α-amino acids.

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SUBSTITUTE SHEET (RULE 26) Nsl table

Amino Acid Reagent (AKP) R-CH-COOH

NH

pK for the COOH-group: 1.8 - 235 pK.dlya NH ₂ rrushsh: -10.7 rKaSNzSOON 8.72: 4.76

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SUBSTITUTE SHEET (RULE 26) Table N ° 2

Indicators of the toxic effect of the reaction masses of the substance after interaction with AKP

There is no clinical picture

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SUBSTITUTE SHEET (RULE 26) Table N ° 3

Indicators of the toxic effect of the hydrolyzate V _x gas and reaction mass after its interaction with AKP

ON about H

Table Ns 4

Data on direct and mutagenic activity of reaction masses after AKP detoxification

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SUBSTITUTE SHEET (RULE 26) Table Ns5

Indicators of the toxic effect of the reaction mass after detoxification of V gas in the process of bio-utilization by microorganisms

There is no clinical picture

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SUBSTITUTE SHEET (RULE 26)

Claims

Claim

1. A method of neutralizing toxic substances, comprising the following steps: the interaction of the toxic substance with a detoxifying reagent; neutralization of the resulting product to pH = 7-8, providing the possibility of further biodegradation; biodegradation to obtain neutralized biomass and wastewater, characterized in that a composition containing at least one natural α-amino acid, a poisonous substance and a detoxifying reagent are taken in a ratio of 1: 1 to 1: 3, and the interaction is carried out at stirring in the temperature range 20-40 ⁰ C for from 0.5 to 1 hour

2. The method according to claim 1, characterized in that mustard gas, lewisite, sarin, soman and Vx hydrolyzate are used as toxic substances.

3. The method according to claims 1 or 2, characterized in that one of the following amino acids is used as the natural α-amino acid: alanine, arginine, aspartic acid, asparagine, valine, histidine, glycine, glutamic acid, glutamine, isoleucine, leucine, lysine, methionine, proline, series, tyrosine, threonine, tryptophan, phenylalanine, cysteine.

4. The method according to p. 1 or 3, characterized in that the product of alkaline hydrolysis of protein-containing industrial waste is used as the amino acid reagent.

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SUBSTITUTE SHEET (RULE 26)