Reaction products of sodium sulphide, sulphur, carbon disulphide and potassium hydroxide

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2017

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

1

Reference substance name:

Reaction products of sodium sulphide, sulphur, carbon disulphide and potassium hydroxide

EC Number:

944-003-3

IUPAC Name:

Reaction products of sodium sulphide, sulphur, carbon disulphide and potassium hydroxide

Test material form:

liquid

Specific details on test material used for the study:

Test Material ID: Reaction products of sodium sulphide, sulphur, carbon disulphide and potassium hydroxide (EC# 944-003-3)
Purity of Test Material: 100% (Note: the test material is placed on the market as 100% solution comprised of 92% water and 8% inorganics that cannot be seperated from the water)
pH of Test Material: >= 12.0 (as reported by the sponsor)
Solubility of Test Material: Soluble (as reported by the sponsor)

Method

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1537

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 98

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 100

Species / strain 5

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

S9 mix (5% v/v)

Test concentrations with justification for top dose:

Final Doses (ug/plate) 0; 1.58; 5.0; 15.8; 25; 50; 158; 500; 5000 with the top dose chosen as the OECD standard limit dose.

Vehicle / solvent:

Vehicle selection was based on existing information when available; if necessary, preliminary solubility trials were to be conducted and documented in the raw data. Generally, the vehicle will be steril water; if the test article is not found to be adequately soluble, other solvents (Such as DMSO) will be considered, followed by aqueous suspending agents (as as 1% methyl cellulose in water) an additional vehicle group may be included when appropriate.

Controls

Controls 1

Positive controls:

yes

Positive control substance:

sodium azide

Controls 2

Positive control substance:

other: ICR 191 Acridine

Details on test system and experimental conditions:

Each of the S. typhimurium and E. coli strains recieved for use on this study were accompanied by documentation that includes lot number, preperation, and expiration dates, and confirmation of phenotype. The tester strains have several features that make them more sensitive for the detection of mutations. The specificity of these strains has provided useful information on the types of mutations that are induced by mutagenic agents (Ames, et al., 1973). The bacterial strains were purchased from Molecular Toxicology, Inc. The initial experiment followed the plate incorporation method, in which the following materials were mixed and poured over the surface of a minimal agar plate: Standard volume of vehicle control, test substance solution, or positive control. 500uL S9 mix or substitution buffer. 100uL bacterial preparation (ST or EC). 2.0 mL overlay agar maintained at approximately 45 deg C.

Rationale for test conditions:

Intermediate and high doses, but not the limit dose, were reduced to account for the high pH of the test substance.

Evaluation criteria:

The results are cinsidered positive (i.e., indicative of mutagenic potential) if: The results for the test item show a substantial increase in revertant colony counts i.e., response MF >/= 2 for strains TA98, TA100, and WP2 uvrA or MF >/= 3 for strains TA1535 and TA1537, with mean value(s) outside the labratory historical control range. Otherwise results were considered negative. The increase must be dose related and/or reproducible, i.e., increases must be obtained at more than one experimental point (at leased one strain, more than one dose level, more than one occasion or with different methodologies).

Statistics:

Means and standard deviations were generated for appropriate quantitave data. Other statistical methods were used if appropriate.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

S. typhimurium TA 1535

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Incomplete lawn at 5000 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Incomplete lawn at 5000 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 3

Key result

Species / strain:

S. typhimurium TA 98

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Incomplete lawn at 5000 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 4

Key result

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Remarks:

Incomplete lawn at 5000 ug/plate

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Test results 5

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

not applicable

Positive controls validity:

valid

Applicant's summary and conclusion

Conclusions:

The Reaction products of sodium sulphide, sulphur, carbon disulphide and potassium hydroxide (EC# 944-003-3) was not mutagenic in bacterial cells, with and without metabolic activation.

Executive summary:

In a bacterial reverse mutation assay (Ames Test) conducted according to OECD Guideline 471 under GLP conditions, the Reaction products of sodium sulphide, sulphur, carbon disulphide and potassium hydroxide (EC# 944-003-3) was not mutagenic, with or without metabolic activation with S9 mix, in any of the bacterial strains tested (TA 1535, TA 1537, TA 98, TA 100, and E. Coli).