Sodium O-ethyl dithiocarbonate

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

There are conclusive but not suffcient data for the classification of substance SEX with regard to mutagenicity/genetic toxicity. It is concluded that the substance SEX does not meet the criteria to be classified for human health hazards for Mutagenicity-Genetic Toxicity

Link to relevant study records

Reference

Reference 1

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

other: published data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment of SEX . In addition, xanthates decompose on aging to form a number of byproducts, depending on the pH, temperature, etc. Risks associated with xanthate are, therefore, a function of the breakdown of the product or un-reacted raw materials remaining in the product.

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EPA OTS 798.5265 (The Salmonella typhimurium Bacterial Reverse Mutation Test)

Deviations:

no

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

S9 mix- rat liver, Aroclor 1254 administered

Test concentrations with justification for top dose:

0.005%, 0.01%, 0.025%, 0.05%, 0.1% v/v

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: none

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

Remarks:

no solvent/vehicle used

Positive controls:

yes

Positive control substance:

other: in the abscence and presence of metabolic activation: 2-aminoanthracene (TA1535), benzo[alpha]pyrene (TA1537, TA100, TA98); in the abscence of metabolic activation: sodium azide (TA1535, TA100), 9-aminoacridine (TA1537), 2-nitrofluorene (TA98);

Remarks:

dicloromethane in a vapour phase (7.5% v/v) was included in each test without metabolic activation

Details on test system and experimental conditions:

METHOD OF APPLICATION: in agar (plate incorporation)

NUMBER OF REPLICATIONS:3/ treatment

DETERMINATION OF CYTOTOXICITY

- Method: abscence or thinning of the background lawn of non-revertant colonies

EXPOSURE TO CS2:
Sets of solidified plates were placed, with lids removed, in stainless steel racks, designed to keep the plates separate and permit atmospheric circulation, inside stainless steel vessels. These vessels were then sealed and partially evacuated. Calculated volumes of carbon disulphide liquid were injected into the vessels via a septum and allowed to vaporize, producing atmospheres containing carbon disulphide at the nominal concentrations
mentioned above.Sterile air was admitted to the vessels in order to equilibrate the contents to atmospheric pressure, and the vessels with their contents were incubated at 37°C for 48 hours. After removal from the vessels, the plates were incubated for a further day in order to permit revertant
colonies to grow to a size large enough to be scored.

Evaluation criteria:

number of revertants/plate

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: yes, only at the highest concentration

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: yes, only at the highest concentration

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: yes, only at the highest concentration

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: yes, only at the highest concentration

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

other: yes, only at the highest concentration

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Carbon disulphide was examined for its mutagenic activity in four histidine-dependent auxotrophs ofSalmonella typhimurium, strains TA98, TA100, TA1535, TA1537. Agar plates, seeded with the tester strains, were exposed to the test material in vapour phase, in the abscence and presence of metabolic activation, in the following nominal concentrations: 0.005%, 0.01%, 0.025%, 0.05% and 0.1% v/v (nominal). These concentrations were selected following preliminary toxicity tests in strain TA 98. Carbon disulphide did not exhibit any mutagenic activity under the conditions of this test. No increases in revertants were obtained in any of the four tester strains following exposure to carbon disulphide at the concentrations tested. Inhibition of bacterial growth, observed as thinning of the background lawn of non-revertant cells and reduction in revertant colony numbers, occurred in all strains with carbon disulphide at a nominal concentration of 0.1 % v/v. The positive and negative controls were valid.
The study was performed according to the OECD Guidelines for Testing of Chemicals No. 471 (1983) and US EPA (TSCA) Guideline § 798.5265 (1985, amended 1987).

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Conclusions:

Interpretation of results :negative

Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment of SEX .
In addition, xanthates decompose on aging to form a number of byproducts, depending on the pH, temperature, etc. Risks associated with xanthate are, therefore, a function of the breakdown of the product or un-reacted raw materials remaining in the product.

Executive summary:

Carbon disulphide was examined for its mutagenic activity in four histidine-dependent auxotrophs of Salmonella typhimurium, strains TA98, TA100, TA1535, TA1537. Agar plates, seeded with the tester strains, were exposed to the test material in vapour phase, in the abscence and presence of metabolic activation, in the following nominal concentrations: 0.005%, 0.01%, 0.025%, 0.05% and 0.1% v/v (nominal). These concentrations were selected following preliminary toxicity tests in strain TA 98. Carbon disulphide did not exhibit any mutagenic activity under the conditions of this test. No increases in revertants were obtained in any of the four tester strains following exposure to carbon disulphide at the concentrations tested. Inhibition of bacterial growth, observed as thinning of the background lawn of non-revertant cells and reduction in revertant colony numbers, occurred in all strains with carbon disulphide at a nominal concentration of 0.1 % v/v. The positive and negative controls were valid. The study was performed according to the OECD Guidelines for Testing of Chemicals No. 471 (1983) and US EPA (TSCA) Guideline § 798.5265 (1985, amended 1987).

Reference 2

Endpoint:

in vitro cytogenicity / chromosome aberration study in mammalian cells

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

other: published data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Qualifier:

according to guideline

Guideline:

OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)

Deviations:

yes

Remarks:

At least duplicate cultures must be used for each experimental point. Only one harvest time was used.

Qualifier:

according to guideline

Guideline:

EPA OPP 84-2

GLP compliance:

yes

Type of assay:

in vitro mammalian chromosome aberration test

Species / strain / cell type:

other: Chinese hamster overy cells

Metabolic activation:

with and without

Metabolic activation system:

S9 mix

Test concentrations with justification for top dose:

3.1, 12.5, 25 ng/ml (without S-9 mix)
125, 500, 1000 ng/ml (with S-9 mix)

Vehicle / solvent:

DMSO

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: Mitomycin C, Cyclophosphamide

Species / strain:

other: Chinese hamster overy cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

 

Dose level [ng/mL]	No. of aberrant cells [%]		Dose level [ng/mL]	No. of aberrant cells [%]
	With MA			Without MA
	Exc. gaps	Inc. gaps		Exc. gaps	Inc. gaps
125	4.5	4.5	3.1	2.0	2.0
500	3.5	4.5	12.5	2.5	2.5
1000	3.5	3.5	25.0	4.0	4.0
Solvent	4.25	4.75	Solvent	2.25	2.25
Positive control	38.5	38.5	Positive control	29.0	29.0

 

Conclusions:

Interpretation of results :negative

No mutagenic activity of Ziram detected.Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Reference 3

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1975

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102

Metabolic activation:

with

Metabolic activation system:

rat liver S9

Test concentrations with justification for top dose:

Not specified for this compound but typically 10, 100, 500, 1000 microgram/plate. (10,000 known also to have been tested).

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

benzo(a)pyrene

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 102

Metabolic activation:

with

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

There were < 70 revertants per 10mg/plate tested.

Remarks on result:

other: all strains/cell types tested

There were < 70 revertants per 10mg/plate tested.

Conclusions:

Interpretation of results :negative

As part of a study of the mutagenic potential of 300 chemicals, ethanol was evaluated in a bacterial reverse mutation (Ames) assay at a plate concentration of up to 10mg/plate in the presence of metabolic activation. There was no evidence of mutagenicity in either of the two strains examined (TA98, TA100). . It should be noted that only two of the normal four bacterial strains were evaluated, so the study cannot be regarded as complete or definitive.
Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Executive summary:

As part of a study of the mutagenic potential of 300 chemicals, ethanol was evaluated in a bacterial reverse mutation (Ames) assay at a plate concentration of up to 10mg/plate in the presence of metabolic activation. There was no evidence of mutagenicity in either of the two strains examined (TA98, TA100). . It should be noted that only two of the normal four bacterial strains were evaluated, so the study cannot be regarded as complete or definitive.

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Reference 4

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Remarks:

no significant deviations noted

Principles of method if other than guideline:

Preincubation as per method of Haworth (Env Mutagen, 5 suppl 1, 3-142, 1983)

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Species / strain / cell type:

S. typhimurium TA 1535

Details on mammalian cell type (if applicable):

Type and identity of media:
- Culture prepared at 37C overnight with shaking in Oxoid #2 broth or in defined minimal medium supplemented with biotin and histidine. Phenotype analysed at time of use.

Species / strain / cell type:

S. typhimurium TA 97

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Culture prepared at 37C overnight with shaking in Oxoid #2 broth or in defined minimal medium supplemented with biotin and histidine. Phenotype analysed at time of use.

Species / strain / cell type:

S. typhimurium TA 98

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Culture prepared at 37C overnight with shaking in Oxoid #2 broth or in defined minimal medium supplemented with biotin and histidine. Phenotype analysed at time of use.

Species / strain / cell type:

S. typhimurium TA 100

Details on mammalian cell type (if applicable):

Type and identity of media:
- Culture prepared at 37C overnight with shaking in Oxoid #2 broth or in defined minimal medium supplemented with biotin and histidine. Phenotype analysed at time of use.

Species / strain / cell type:

S. typhimurium, other: TA104

Details on mammalian cell type (if applicable):

- Type and identity of media:
- Culture prepared at 37C overnight with shaking in Oxoid #2 broth or in defined minimal medium supplemented with biotin and histidine. Phenotype analysed at time of use.

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254 induced male SD rat at 10 and 30% and male Syrian hamster liver S9 fraction, at 10 and 30%.

Test concentrations with justification for top dose:

1; (3; 10; 33;)100; 333; 1,000; 3,333; 10,000 microgram/plate. Concentrations in brackets only tested with TA97 using 30% hamster S9.

Vehicle / solvent:

water

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

Remarks:

water

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene, used for all strains with metabolic activation

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 4-nitro-o-phenylenediamine, used for TA98 without metabolic activation

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

Remarks:

used for TA97 without metabolic activation.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

sodium azide

Remarks:

used for TA1535 and T100 without metabolic activation.

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

methylmethanesulfonate

Remarks:

used for TA104 without metabolic activation.

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- Preincubation period: 20 mins at 37C
- Expression time (cells in growth medium): 2 days at 37C

NUMBER OF REPLICATIONS: 5 plus complete repeat of experiment.

DETERMINATION OF CYTOTOXICITY
- Dose range for main test assessed using TA100. Toxic concentrations defined as those producing a decrease in the background number of his+ colonies and/or a clearing in the background lawn. If no toxicity was seen, the maximum dose tested was 10000ug/plate

Evaluation criteria
Only considered non mutagenic if negative in all strains with and without activation and with both S9 extracts at both concentrations

Evaluation criteria:

Combination of magnitude of increase in number of his+ revertants and shape of dose-response curve.

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium TA 97

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Species / strain:

S. typhimurium, other: TA104

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not determined

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- None

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Dose-effected related observations: Ethanol at any dose did not produce a 2-fold increase in his+ revertants in the absence or presence of rat or hamster liver extracts.

Conclusions:

Interpretation of results :negative

Ethanol failed to induce reversions in any S. typhimurium tester strain with or without metabolic activation over a wide range of doses up to 10 mg/plate.Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Executive summary:

In a reverse mutation assay in bacteria strains TA97, TA98, TA100, TA104 and TA1535 there was no evidence of mutation up to a maximum plate concentration of 10mg/plate with and without metabolic activation systems derived from two species (rat and hamster) used at two different concentrations.

Reference 5

Endpoint:

in vitro gene mutation study in mammalian cells

Remarks:

Type of genotoxicity: gene mutation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1988

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)

Deviations:

no

Remarks:

no significant deviations noted

Principles of method if other than guideline:

Method: other: Clive et al. (Muta Res, 59, 61, 1979) with some minor modifications to reduce experiment time and plating efficiency. Study examined a large number of chemicals.

GLP compliance:

not specified

Type of assay:

mammalian cell gene mutation assay

Target gene:

TK forward mutation

Species / strain / cell type:

mouse lymphoma L5178Y cells

Details on mammalian cell type (if applicable):

- Type and identity of media: Fisher's medium with 10% horse serum, adjusted to pH 7.2
- Source: Boroughs Welcome Co, Research Triangle Park, NC, USA
- Periodically "cleansed" against high spontaneous background: yes by treatement with thymidine, hypoxanthine, methotrexate and glycine

Additional strain / cell type characteristics:

not specified

Metabolic activation:

with and without

Metabolic activation system:

Liver S9 from Sprague-Dawley rat livers, animals pre-treated with Aroclor 1254.

Test concentrations with justification for top dose:

0.092, 0.184, 0.369, 0.553, 0.738 mol/l without activation; 0.414, 0.465 and 0.517 mol/l with activation

Untreated negative controls:

yes

Negative solvent / vehicle controls:

not specified

True negative controls:

no

Positive controls:

yes

Positive control substance:

4-nitroquinoline-N-oxide

2-nitrofluorene

benzo(a)pyrene

cyclophosphamide

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium

DURATION
- Exposure duration: 4 hours
- Expression time (cells in growth medium): 48 hours
- Selection time (if incubation with a selection agent): no data
- Fixation time (start of exposure up to fixation or harvest of cells):

SELECTION AGENT (mutation assays): trifluorothymidine

NUMBER OF REPLICATIONS: 3 per dose level but 6 for negative control.

DETERMINATION OF CYTOTOXICITY
- Mitotic index: Not strictly applicable. Total growth cf. controls were 88, 84, 53, 34 and 17% from lowest to highest concentrations in the absence of activation. With activation, total growth was 43, 24, and 6% from lowest to highest concentration.

Evaluation criteria:

Two fold or greater increase in mutation frequency at 10% or greater total growth cf. controls.

Statistics:

Tested for normal distribution and then analysis of variance and 2-tailed Student's t-test against controls.

Species / strain:

mouse lymphoma L5178Y cells

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not examined

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- None

ADDITIONAL INFORMATION ON CYTOTOXICITY: see table below
- Frequency of reversions etc: Without activation, mutation index values from lowest to highest dose were 1.3, 1.1, 1.2, 1.1 and 1.6. With metabolic activation these values were 1.1, 1.3 and 1.8.
- Dose-effect related observations: No dose-effect related observations were seen.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Only at the maximum concentration, with metabolic activation was total growth <10% control. Without activation, the lowest and highest concentrations of ethanol produced statistically significant increases in mutation frequency.

Without metabolic activation

Concentration (mols/litre)	Total growth	Mutation frequency	Mutation index
0	100%	80, 99	1.0
0.0922	88%	118*	1.3
0.184	84%	94	1.1
0.369	53%	104	1.2
0.553	34%	101	1.1
0.738	17%	140**	1.6

With metabolic activation

Concentration (mols/litre)	Total growth	Mutation frequency	Mutation index
0	100%	63, 46	1.0
0.414	43%	62	1.
0.465	24%	70	1.3
0.517	6%	97**	1.8

*significant, ** highly significant

Rates of spontaneous mutation frequencies in study: without metabolic activation 76 +/-25, with metabolic activation 86 +/-33.

Results are supported by those of Amacher, D., et al. (1980) Mutat. Res. 72:447 -474

 

Conclusions:

Interpretation of results :negative

Ethanol is judged not to have significant mutagenic activity in this system.
Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Executive summary:

In a mammalian cell mutation study using mouse lymphoma lymphoma cells in the TK forward mutation assay, ethanol was found to be non mutagenic with and without metabolic activation at very high doses up to and including those that cause significant cytotoxicity (typically in the region 0.3 -0.5M.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Link to relevant study records

Reference

Reference 1

Endpoint:

in vivo mammalian germ cell study: cytogenicity / chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

other: published data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Qualifier:

according to guideline

Guideline:

OECD Guideline 483 (Mammalian Spermatogonial Chromosome Aberration Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.23 (Mammalian Spermatogonial Chromosome Aberration Test)

Qualifier:

according to guideline

Guideline:

EPA OPP 84-2

GLP compliance:

yes

Type of assay:

mammalian germ cell cytogenetic assay

Species:

mouse

Strain:

NMRI

Sex:

male

Details on test animals or test system and environmental conditions:

- Source: Charles River, Germany
- Age at study initiation: min. 10 weeks
- Weight at study initiation: ca. 30 g

Route of administration:

oral: gavage

Vehicle:

0.5% carboxymethylcellulose

Duration of treatment / exposure:

6, 24 and 48 h

Frequency of treatment:

Single application

Post exposure period:

n.a.

Remarks:

Doses / Concentrations:
20, 67 and 200 mg/kg b.w.
Basis:
nominal conc.

No. of animals per sex per dose:

5 males (positive control only 4)

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide (CPA)
- Dose: 140 mg/kg b.w.

Tissues and cell types examined:

Spermatogonia

Details of tissue and slide preparation:

In the test article and the negative control groups 100 well spread metaphases per animal were scored for cytogenetic damage on coded slides. In the positive control group 50 metaphases per animal were scored. Only metaphases with the characteristic chromosome number of 40 were included in the analysis. To describe a cytotoxic effect the mitotic index (% cells in mitosis) was determined.
Five animals per test group were evaluated as described. The remaining animals of each test group were evaluated in case animals died in its test group.

Sex:

male

Genotoxicity:

negative

Toxicity:

yes

Remarks:

Tested in pre-experiments.

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Group	Dose [mg/kg]	Exposure [h]	No. of cells scored	Aberrant cells [%]		Mitotic index [%]
				Incl. gaps	Excl. gaps
Ziram	200	6	500	0.0	0.0	1.40
Vehicle control	0	24	500	0.4	0.2	2.80
Ziram	20	24	500	0.0	0.0	1.40
Ziram	67	24	500	0.2	0.0	2.34
Ziram	200	24	500	0.2	0.2	1.84
Positive control	140	24	200	8.0	7.5	0.34
Ziram	200	48	500	0.8	0.4	1.16

Conclusions:

Interpretation of results : negative
No mutagenic activity of Ziram detected. Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Reference 2

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

other: published data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Qualifier:

according to guideline

Guideline:

OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.11 (Mutagenicity - In Vivo Mammalian Bone-Marrow Chromosome Aberration Test)

GLP compliance:

yes

Type of assay:

chromosome aberration assay

Species:

mouse

Strain:

NMRI

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: BRL Tierfarm Füllinsdorf, Switzerland
- Age at study initiation: min. 10 weeks
- Weight at study initiation: ca. 30 g

Route of administration:

oral: gavage

Vehicle:

Carboxymethylcellulose

Duration of treatment / exposure:

6, 24 and 48 h

Frequency of treatment:

Single application

Post exposure period:

n.a.

Remarks:

Doses / Concentrations:
0, 40, 120 and 400 mg/kg b.w.
Basis:
nominal conc.

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Positive control(s):

Cyclophosphamide (CPA)
- Dose: 20 mg/kg b.w.

Tissues and cell types examined:

Bone marrow cells

Details of tissue and slide preparation:

At least 50 well spread metaphases per animal were scored for cytogenetic damage on coded slides. Only metaphases with the characteristic chromosome number of 40 were included in the analysis. To describe a cytotoxic effect the mitotic index (% cells in mitosis; 1000 cells are scored) was determined.
Five animals per sex and group were evaluated as described. The remaining animal of each test group was evaluated in case an animal died in its test group spontaneously or due to gavage error.

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Remarks:

Tested in pre-experiments.

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

valid

Group	Dose [mg/kg]	Exposure [h]	No. of cells scored	Aberrant cells [%]		Mitotic index [%]
				Incl. gaps	Excl. gaps
Ziram	400	6	500	0.4	0.2	3.54
Vehicle control	0	24	500	0.4	0.0	4.91
Ziram	40	24	500	1.0	0.6	4.03
Ziram	120	24	500	0.6	0.0	5.50
Ziram	400	24	500	1.6	1.2	3.52
Positive control	20	24	500	28.4	27.8	5.00
Ziram	400	48	500	0.2	0.2	5.08

Conclusions:

Interpretation of results : negative
No mutagenic activity of Ziram detected. Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Reference 3

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

other: published data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

No positive control.

Qualifier:

according to guideline

Guideline:

EU Method B.12 (Mutagenicity - In Vivo Mammalian Erythrocyte Micronucleus Test)

Qualifier:

according to guideline

Guideline:

EPA OPP 84-2

GLP compliance:

yes

Type of assay:

micronucleus assay

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Source: Charles River, USA
- Age at study initiation: 5-6 weeks
- Weight at study initiation: no data

Route of administration:

oral: feed

Duration of treatment / exposure:

89 days

Frequency of treatment:

daily

Post exposure period:

none

Remarks:

Doses / Concentrations:
0, 25, 75, 225 and 675 ppm
Basis:
nominal in diet

No. of animals per sex per dose:

50

Control animals:

yes, plain diet

Tissues and cell types examined:

Peripheral blood

Details of tissue and slide preparation:

Peripheral blood smears were prepared from 5 male and 5 female animals in each group. The smears were stained using Giemsa then examined by light microscopy for the presence of micronuclei in normochromatic and polychromatic erythrocytes (1000 cells of each type were examined from each animal). The ratio of polychromatic to normochromatic erythrocytes was assessed by examination of at least 1000 erythrocytes from each animal.

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

valid

Negative controls validity:

not examined

Positive controls validity:

not examined

Dose level [ppm]	Ration p/n (mean)	Incidence mnp (mean)	Incidence mnn (mean)
Control	0.108	0.9	1.1
25*	0.143	0.6	1.7
75**	0.124	1.2	2.0
225	0.095	0.8	0.7
675	0.135	0.5	1.2

p/n Ratio of polychromatic to normochromatic erythrocytes

mnp No. of micronucleated cells observed per 1000 polychromatic erythrocytes

mnn No. of micronucleated cells observed per 1000 normochromatic erythrocytes

* Prepared at 29 ppm initially to allow for loss during storage

** Prepared at 83 ppm initially to allow for loss during storage

 

Conclusions:

Interpretation of results : negative
No mutagenic activity of Ziram detected. Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

Reference 4

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

other: published data

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment of SEX .

Qualifier:

according to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

GLP compliance:

not specified

Type of assay:

micronucleus assay

Species:

mouse

Strain:

CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories (UK)
- Age at study initiation: 4-5 weeks
- Weight at study initiation: 16.1-26 g
- Assigned to test groups randomly: yes
- Housing: high density polypropylene cages with stainless steel taps
- Diet: ad libitum
- Water: supplied via a polythene bottle and sipper tube
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21±2
- Humidity (%): 55 ±15 %
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light):12

Route of administration:

inhalation: vapour

Vehicle:

no vehicle used

Details on exposure:

TYPE OF INHALATION EXPOSURE: snout only

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
-Animals were exposed to the test material by snout-only inhalation. Prior to exposure of animals, the test material atmospheres were generated for each exposure chamber and samples analysed. Each mouse was placed in an individual polymethyl methacrylate restraining tube so that only the snout protruded. Each restraining tube was marked with the animal and group numbers. The restraining tubes were attached to the appropriate chamber so that the snout of each mouse projected into the lumen of the chamber. When the pre-exposure observations were complete, the syringe pump was switched on and the exposure timed for six hours following a 4.5 minute equilibration period, the theoretical time required for the concentration of vapour to reach 90% of its final value under the conditions of exposure employed (Silver and Arsenal, 1946). After six hours, the test atmosphere
supply was switched off and the mice removed from the restraining tubes for examination.

Duration of treatment / exposure:

6 h

Frequency of treatment:

once

Remarks:

Doses / Concentrations:
0, 467, 1558, 4675 mg/m3 (150, 500, 1500 ppm)
Basis:
nominal conc.

No. of animals per sex per dose:

10 (5 for positive control)

Control animals:

yes

Positive control(s):

chlorambucil
- Route of administration: oral
- Doses / concentrations: 30 mg/kg

Tissues and cell types examined:

bone marrow erythrocytes

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION: based on the preliminary toxicity testing

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields): samples were taken 24 and 48 h after treatment

DETAILS OF SLIDE PREPARATION: Animals were killed by cervical dislocation following carbon dioxide inhalation. Femurs from each animal were rapidly dissected out and cleaned of adherent tissue. The epiphyses were cut off to obtain access to the marrow canal. Marrow cells were flushed out with 2.5 ml foetal calf serum using a syringe and needle. The recovered cells were centrifuged at 1000 rpm for five minutes. The bulk of the supernatant fluid was discarded and the cell pellet resuspended in the remaining fluid. Single drops of the cell suspension were transferred to clean, dry slides, two or three smears (for the preliminary toxicity test or main micronucleus test respectively) prepared, and the slides left to air-dry. Following fixation in methanol for ten minutes, they were stained manually in 5% Giemsa stain (in Sorensen's buffer: pH 6.8), washed in buffer, air-dried, cleared for five minutes in xylene and made permanent using DPX mountant.


METHOD OF ANALYSIS: The slides were examined under the light microscope. At high magnification (x 1000, oil immersion) a total of at least 2000 erythrocytes per animal were examined. Each erythrocyte scored was classed as polychromatic or mature. Each erythrocyte scored was also examined for the presence or absence of micronuclei. Thereafter, the frequencies of micronucleated cells per 1000 erythrocytes were calculated. The ratio of polychromatic to mature cells was also determined (indicating the rythm of cell division). The frequency of micronuclei in polychromatic cells provides an index of induced genetic damage.

Evaluation criteria:

Positive for clastogenicity was a statistically and biologically significant increase in micronucleated polychromatic cells, compared to vehicle control, in at least one treatment group; particularly if supported by evidence of a dose-related response.

Statistics:

Mann-Whitney U procedure (Mann and Whitney, 1942), two-tailed test, one-tailed test.

Sex:

male/female

Genotoxicity:

negative

Toxicity:

no effects

Vehicle controls validity:

not examined

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDY
- Dose range: 45, 150, 450, 1500, 2000 ppm
- Clinical signs of toxicity in test animals: unconscious and death at 2000 ppm, unconscious, slow and laboured respiration, all extremities red coloured. No adverse reactions to treatment were observed in animals exposed to 450, 150, 45 ppm test substance.
- Evidence of cytotoxicity in tissue analyzed: no bone marrow toxicity observed
- Harvest times: 48 hours

RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): see table
- Ratio of PCE/NCE (for Micronucleus assay): see table

The micronucleus test - group means and standard deviations (sd) by sex

test group	dose (ppm)	sampling time (h)	Sex	total polychromatic cells scored	total micronucleated polychromatic cells	mean micronucleated polychromatic cells per 1000 and sd	total mature cells scored	total micronucleated mature cells per 1000 and sd	polychromatic cells/mature cells
Air	0	24	M	5183	7	1.3±0.9	5652	4	0.7±0.4
			F	5709	3	0.5±0.5	5607	3	0.6±0.9
Carbon disulphide	150		M	5208	2	0.4±0.5	5871	4	0.6±0.9
			F	6030	6	1.0±0.7	5469	5	0.9±0.8
	500		M	6194	14	2.0±1.6	5505	2	0.4±0.5
			F	5788	10	1.6±1.1	5148	3	0.6±0.5
	1500		M	6713	8	1.0±1.2	5089	2	0.4±0.5
			F	7640	9	1.2±0.9	5129	2	0.4±0.5
Chlorambucil	30 mg/kg		M	5473	322	59.0±27.6	5176	3	0.6±0.9
			F	5807	403	69.3±27.5	5201	7	1.3±0.8
Air	0	48	M	5090	4	0.8±0.8	6101	7	1.1±0.8
			F	5588	4	0.7±0.8	4561	1	0.2±0.4
Carbon disulphide	150		M	5353	2	0.4±0.5	5699	4	0.7±0.6
			F	5976	9	1.5±1.1	5079	0	0.0±0.0
	500		M	6153	7	1.1±1.2	5300	3	0.6±0.9
			F	6518	6	0.9±0.6	5105	2	0.4±0.5
	1500		M	7324	8	1.0±0.9	5072	0	0.0±0.0
			F	6558	15	2.1±1.4	5022	0	0.0±0.0

 

Conclusions:

Interpretation of results : negative
No evidence of induced chromosomal or other damage leading to the formation of micronuclei in erythrocytes of the bone marrow was detected after exposure of the animals to CS2 via inhalation.
Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment off SEX .
In addition, xanthates decompose on aging to form a number of byproducts, depending on the pH, temperature, etc. Risks associated with xanthate are, therefore, a function of the breakdown of the product or un-reacted raw materials remaining in the product.

Executive summary:

The effect of carbon disulphide on chromosome structure in the bone marrow erythrocytes of mice was examined. The animals (males and females) were exposed via inhalation snout-only, for 6 h to the following concentrations: 0, 467, 1558, 4675 mg/m3 (0, 150, 500, 1500 ppm). The exposure concentrations were based on a preliminary toxicity test. Chlorambucil (30 mg/kg bw) was used as a positive control, adminstered via the oral route. Animals were sacrifised and examined 24 and 48 h after exposure. No evidence of induced chromosomal or other damage leading to the formation of micronuclei in erythrocytes of the bone marrow was detected, uder the present test condition, after exposure of the animals to CS2 via inhalation. Mice exposed at 1500 ppm, however, showed a small increase in the ratio of polychromatic/mature cells, which may indicate disturbance of erythropoiesis. Carbon disulphide was tested for induction of micronuclei in the bone marrow ertythrocytes of mice according to the OECD Guidelines 474 (1983).

Reference 5

Endpoint:

genetic toxicity in vivo, other

Remarks:

Type of genotoxicity: other: review paper covering many studies

Type of information:

other: published data

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment of SEX . In addition, xanthates decompose on aging to form a number of byproducts, depending on the pH, temperature, etc. Risks associated with xanthate are, therefore, a function of the breakdown of the product or un-reacted raw materials remaining in the product.

Qualifier:

no guideline available

Principles of method if other than guideline:

Review of a large series of publications on studies in which a wide array of methods was applied.

GLP compliance:

not specified

Type of assay:

other: review paper covering many studies

Species:

other: A series of studies with various animal species was reviewed.

Strain:

other: A series of studies with various animal species was reviewed.

Sex:

male/female

Details on test animals or test system and environmental conditions:

Not applicable: review paper.

Route of administration:

other: Invasive (injection) and non-invasive (inhalation, oral, dermal) routes were employed in the different studies reviewed.

Vehicle:

Not applicable: review paper.

Details on exposure:

Not applicable: review paper.

Duration of treatment / exposure:

Not applicable: review paper.

Frequency of treatment:

Not applicable: review paper.

Conclusions:

Interpretation of results : negative
In the majority of the in vivo tests assessing CS2 mutagenic potential, a negative result was obtained, except for one case. The validity of the studies is uncertain due to technical issues (e.g. invalid controls).
Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. Reaction mass of SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment of Reaction mass of SEX .
In addition, xanthates decompose on aging to form a number of byproducts, depending on the pH, temperature, etc. Risks associated with xanthate are, therefore, a function of the breakdown of the product or un-reacted raw materials remaining in the product.

Executive summary:

In male and female rats inhaling 63 or 125 mg carbon disulfide/m3, 7 hours per day for 1 or 5 days, there was no significant increase in the frequency of chromosomal aberrations in bone marrow cells (Belisles et al., 1980). In another study (Vasil’eva, 1982), oral exposure to carbon disulfide gave a mutagenic response, manifested as chromosomal aberrations and polyploid cells in the bone marrow of female rats and in rat embryos exposed on days 10–13 of gestation. According to the reviewer,'it is difficult to assess the validity of these findings, as the reporting was brief e.g., the statistical significance was often not indicated and the effective dose was not reported, except to indicate that it was one-tenth of the LD50 '. In the investigation of Belisles et al. (1980), male rats were exposed to 63–125 mg/m3 of CS2, 7 h/d for 5 d; no significant increase in dominant lethal mutations was observed, nor was there a dose related increase in sperm abnormalities in rats or mice exposed according to the same protocol. However, lack of an effect on sperm abnormalities in positive control rats suggests that there was a problem with the test methods in this study.

Reference 6

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1993

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)

Deviations:

yes

Remarks:

No positive control, no rationale for dose or duration of exposure provided, low number of animals per group

Principles of method if other than guideline:

Method: other: Preparation and analysis of pulmonary alveolar macrophages described below.

GLP compliance:

not specified

Type of assay:

micronucleus assay

Species:

rat

Strain:

other: BD6

Sex:

male

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Weight at study initiation: 180 - 200g
- Diet: Standard rodent diet
- Water: water ad libitum alone or with added ethanol.
- Acclimation: 1 week

Route of administration:

oral: drinking water

Vehicle:

drinking water

Duration of treatment / exposure:

Exp 1: 10 days (5% or 10% ethanol)
Exp 2: 30 days (5% ethanol)
Exp 3: 23 days (5% ethanol)

Remarks:

Doses / Concentrations:
5% or 10%
Basis:
nominal in water

No. of animals per sex per dose:

51 in 3 separate experiments. 3 animals per group.

Control animals:

yes, concurrent vehicle

Tissues and cell types examined:

At the end of the treatments animals were killed and bronchoalveolar lavage was performed with NaCl solution via a cannula inserted in the trachea. Cells were then washed twice and centrifuged, fixed with methanol and then stained with Giemsa solution.
Bone marrow cells were recovered by means of the paint-brush technique (Albanese et al. Mutat. Res, 182, 323-332, 1987). Cells were stained with Mayer's Haemalum, rinsed with water and further stained with eosin. Slides were dipped in xylene and mounted in Eukitt.

Slides were blind-coded and assessed by two readers, each one examining at least 2000 PAM (12000 cells scored per experimental group), and 1000 PCE per rat (6000 cells scored per experimental group ).

Statistics:

Student's t-test

Sex:

male

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Additional information on results:

No effect on micronucleus incidence was observed.
Binucleated and polynucleated pulmonary alveolar macrophages were significantly enhanced by ethanol treatment.
10% dose significantly decreased the PCE/NCE ratio and was cytotoxic to bone marrow.
Assuming a rat water consumption of 50ml/kg/day 5% ethanol in drinking water (assume v/v) is equivalent to a dose of 2.0g/kg and 10% to 3.9g/kg.

No effect on micronucleus incidence was observed. Binucleated and polynucleated pulmonary alveolar macrophages were significantly enhanced by ethanol treatment. 10% dose significantly decreased the PCE/NCE ratio and was cytotoxic to bone marrow. Assuming a rat water consumption of 50ml/kg/day 5% ethanol in drinking water (assume v/v) is equivalent to a dose of 2.0g/kg and 10% t

Conclusions:

Interpretation of results: negative
In an in vivo micronucleus study, male rats were exposed to 5% v/v or 10% ethanol in drinking water for a period of 10 days, 23 days or 30 days in three separate experiments. These were equivalent to doses of 2.0 and 3.9g/kg. The treatment did not induce any changes in the frequency of micronucleated polychromatic erythrocytes and pulmonary alveolar macrophages. However, ethanol significantly enhanced the number of binucleated and polynucleated pulmonary alveolar macrophages, possibly reflecting some generic disturbances in cell cycle control and nuclear-cytoplasmic balance although this is not clearly defined. A dose of 5% for 10 days is within the guideline but the other doses were about 2x those normally recommended (on a dose/time basis).
Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate and sodium hydroxide. Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Executive summary:

In an in vivo micronucleus study, male rats were exposed to 5% v/v or 10% ethanol in drinking water for a period of 10 days, 23 days or 30 days in three separate experiments. These were equivalent to doses of 2.0 and 3.9g/kg. The treatment did not induce any changes in the frequency of micronucleated polychromatic erythrocytes and pulmonary alveolar macrophages. However, ethanol significantly enhanced the number of binucleated and polynucleated pulmonary alveolar macrophages, possibly reflecting some generic disturbances in cell cycle control and nuclear-cytoplasmic balance although this is not clearly defined. A dose of 5% for 10 days is within the guideline but the other doses were about 2x those normally recommended (on a dose/time basis).

Reference 7

Endpoint:

in vivo mammalian somatic cell study: cytogenicity / bone marrow chromosome aberration

Remarks:

Type of genotoxicity: chromosome aberration

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

1981

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 475 (Mammalian Bone Marrow Chromosome Aberration Test)

Deviations:

yes

Remarks:

No positive control, no rationale on dose selection provided, number of animals in control group low, cell sampling 2.5hr after colcemid instead of 4-5hrs.

GLP compliance:

not specified

Type of assay:

chromosome aberration assay

Species:

hamster, Chinese

Strain:

not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 10-20 weeks.
- Diet: (Altromin 7024)
- Water: Drinking water ad libitum. Controls received plain water.

Route of administration:

oral: drinking water

Vehicle:

Vehicle(s)/solvent(s) used: water.

Duration of treatment / exposure:

12 week

Remarks:

Doses / Concentrations:
10% in the drinking water during week 1, 15% in weeks 2 - 3, 20% in weeks 4 - 12.
Basis:
nominal in water

No. of animals per sex per dose:

Ethanol treated 8 females, 10 males.
Controls 4 females, 4 males.

Control animals:

yes, concurrent vehicle

Tissues and cell types examined:

CA: At least 100 metaphases analysed for each animal. Animals received 2mg/kg colcemide ip prior to sacrifice.
SCE: Animals were implanted tablets containing BrdU subcutaneously and received 2mg/kg colcemide ip prior to sacrifice.

Sex:

male/female

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

valid

Negative controls validity:

not applicable

Positive controls validity:

not applicable

Additional information on results:

2.3% aberrant metaphases were detected in controls (900 metaphases analysed), and 3.7 % in treated animals (400 metaphases analysed). No sex difference was evident. Mitotic index was significantly elevated in ethanol treated group (4.2%) compared to the control group (2.7%) (p<0.001).
There were no differences in the frequency of SCE per metaphase analysed between ethanol group (3.86 +/-0.12; 410 metaphases anlaysed) and control group (3.75 +/-0.15; 210 metaphases analysed).

2.3% aberrant metaphases were detected in controls (900 metaphases analysed), and 3.7 % in treated animals (400 metaphases analysed). No sex difference was evident. Mitotic index was significantly elevated in ethanol treated group (4.2%) compared to the control group (2.7%) (p<0.001).

There were no differences in the frequency of SCE per metaphase analysed between ethanol group (3.86 +/-0.12; 410 metaphases anlaysed) and control group (3.75 +/-0.15; 210 metaphases analysed).

Conclusions:

Interpretation of results : negative
In a chromosome abberation study, male and female Chinese hamsters were administered ethanol in drinking water at 10% v/v during the first week, 15% v/v during the 2nd and 3rd, and 20% v/v for additional 8 weeks. Dosing was equivalent for the latter two thirds of the study of 31g/kg for males and 37g/kg for females. Analysis of bone marrow cells did not show any increase in chromosomal aberrations and sister chromatic exchanges in the ethanol group compared with the vehicle control group and doses well in excess of what is normally recommended in the guideline.
Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate and sodium hydroxide. Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

Executive summary:

In a chromosome abberation study, male and female Chinese hamsters were administered ethanol in drinking water at 10% v/v during the first week, 15% v/v during the 2nd and 3rd, and 20% v/v for additional 8 weeks. Dosing was equivalent for the latter two thirds of the study of 31g/kg for males and 37g/kg for females. Analysis of bone marrow cells did not show any increase in chromosomal aberrations and sister chromatic exchanges in the ethanol group compared with the vehicle control group and doses well in excess of what is normally recommended in the guideline.

Ethyl Alcohol is both reagents used in the manufacture, as well as decomposition products of xanthates. Therefore, the health effects of Ethyl Alcohol need to be considered in the assessment of sodium ethyl xanthate.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Additional information

Additional information from genetic toxicity in vitro:

No mutagenic activity of CS2 detected in the reliable study of Akzo Chemicals International BV 1992.Carbon disulphide was examined for its mutagenic activity in four histidine-dependent auxotrophs of Salmonella typhimurium, strains TA98, TA100, TA1535, TA1537.Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment of SEX .

No evidence of induced chromosomal or other damage leading to the formation of micronuclei in erythrocytes of the bone marrow was detected after exposure of the mouse to CS2 via inhalationin the reliable study of Akzo Chemicals International BV 1992..

Carbon disulphide is both a reagent in the manufacture, as well as a decomposition product of xanthates. SEX readily decomposes to carbon disulphide, especially in the presence of moisture/water. Therefore, the health effects of carbon disulphide (CS2) need to be considered in the assessment of SEX .

No mutagenic activity of Ethyl Alcohol detected in the reliable study of Zeiger, E., Anderson, B., Haworth, S., Lawlor, T, Mortelamns, K,1992.Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

 

No mutagenic activity of Ethyl Alcohol detected in the reliable study ofWangenheim, J. and Bolcsfoldi, G.1988.In a mammalian cell mutation study using mouse lymphoma lymphoma cells in the TK forward mutation assay, ethanol was found to be non mutagenic with and without metabolic activation at very high doses up to and including those that cause significant cytotoxicity (typically in the region 0.3 -0.5M.

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

 

No mutagenic activity of Ethyl Alcohol detected in the reliable study of McCann, J., Choi, E., Yamasaki, E., Ames, B.N.1975.As part of a study of the mutagenic potential of 300 chemicals, ethanol was evaluated in a bacterial reverse mutation () assay at a plate concentration of up to 10 mg/plate in the presence of metabolic activation. There was no evidence of mutagenicity in either of the two strains examined (TA98, TA100). . It should be noted that only two of the normal four bacterial strains were evaluated, so the study cannot be regarded as complete or definitive.

Ethanol/ Ethyl Alcohol is both reagents used in the manufacture of sodium O-ethyl dithiocarbonate . Therefore, Ethanol/ Ethyl Alcohol need to be considered in the assessment of sodium O-ethyl dithiocarbonate

No mutagenic activity of Ziram detectedin the reliable study of Brooker, P.C. & Akhurst, L.C.1989.

Dithiocarbamates are related compounds to xanthates. This is organosulfur compound is obtained by treating carbon disulfide with amine in the presence of sodium or potassium hydroxide: They arise from the reaction of the amine with CS2.

 

 

Justification for selection of genetic toxicity endpoint
Negative in all test conducted.

Justification for classification or non-classification

Based on the hazard assessment of SEX in section 2.1 and 2.2. in IUCLID 6., available data for the substance and following the “Guidance on InformationRequirement and Chemical Safety Assessment R.8. Characterisation of dose [concentration]- response for human health” andaccording to the criteria described in Directive 67/548 and in the CLP Regulation:

 

Directive 67/548	Mutagenicity-Genetic Toxicity Muta. Cat. 1; R46 May cause heritable genetic damage. Muta. Cat. 2; R46 May cause heritable genetic damage. Muta. Cat. 3; R68 Possible risk of irreversible effects.
CLP	Germ cell mutagenicity Muta. 1A Muta. 1B Muta. 2 H340: May cause genetic defects <state route of exposure if it is conclusively proven that no other routes of exposure cause the hazard>. H341: Suspected of causing genetic defects <state route of exposure if it is conclusively proven that no other routes of exposure cause the hazard>.

 

It is concluded that the substance SEX does not meet the criteria to be classified for human health hazards for Mutagenicity-Genetic Toxicity