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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Study period:
June 20, 2014 -
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: The study was conducted according to GLP and in compliance with OECD guideline 471.

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2014

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Reference substance name:
Not applicable
EC Number:
940-936-5
Molecular formula:
C4H12NCl.C4H12NOCl
IUPAC Name:
Not applicable
Test material form:
other: Halo Salt was prepared as a formulation in dimethylsulfoxide (DMSO) at a concentration of 50 mg/mL.
Details on test material:
- Name of test material (as cited in study report): Halo Salt
- Physical state: liquid
- Analytical purity: 100%
- Impurities (identity and concentrations): Al, Ca,Cr,Co,Cu,Fe,Pb,Mg,Ni,K,Na,Sn,Ti,Zn < 100 ppb
- Composition of test material, percentage of components: Tetramethylammonium Hydroxide 1.34 (wt.%), Available Chlorine 8.31 (wt.%), Stabilizer 0.12 (wt%)
- Lot/batch No.: 0000066680
- Storage condition of test material: Store in well-closed, light-resistant containers

Method

Target gene:
Histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
other: The Salmonella tester strains have a deep rough mutation (rfa-) that causes the cell wall to be defective in the lipopolysaccharide coat resulting in increased permeability to large molecules.
Species / strain / cell type:
E. coli WP2 uvr A
Additional strain / cell type characteristics:
other: All tester strains have a DNA deletion that affects repair of ultraviolet light damage which greatly increases sensitivity in detecting mutagens.
Metabolic activation:
with and without
Metabolic activation system:
Rat liver S9 fraction
Test concentrations with justification for top dose:
The initial assay:
Halo Salt was tested at 25, 50, 100, 250, 500, 1000, 2500 and 5000 µg/plate using the plate incorporation method

The confirmatory assay:
Halo Salt was tested at 100, 250, 500, 1000, 2500 and 5000 µg/plate using the preincubation method.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
Controls
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
Positive controls:
yes
Positive control substance:
4-nitroquinoline-N-oxide
2-nitrofluorene
sodium azide
other: 2-aminoanthracene, ICR-191 acridine
Details on test system and experimental conditions:
The study was conducted according to the OECD guideline 471.
Evaluation criteria:
Criteria for Positive Response:
The test item was considered positive for mutagenicity if it induced an increase of revertants per plate with increasing concentration. The increases should be at least two times the vehicle control background frequency for strains with high spontaneous levels (i.e., TA100) and three times for those with low spontaneous levels (TA1537, TA98, TA1535 and WP2 uvrA). These increases should be seen in at least two or more successive concentrations or the response should be repeatable at a single concentration.

Criteria for Negative Response:
The test item was considered to be negative for inducing mutagenicity if it did not induce a response which fulfills the criteria for a positive response.

Criteria for Equivocal Response
Cases which did not clearly fit into the positive or negative criteria may be judged equivocal. In these cases the Study Director, based on sound scientific judgment, may take additional factors into consideration in evaluating the test results.
Statistics:
For each concentration level and for each condition, the mean revertant count and standard deviation (SD) were determined.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
The concentrations tested in the confirmatory assay were 100, 250, 500, 1000, 2500 and 5000 µg/plate using the preincubation method.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Species / strain:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Remarks:
The concentrations tested in the confirmatory assay were 100, 250, 500, 1000, 2500 and 5000 µg/plate using the preincubation method.
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative with metabolic activation
negative without metabolic activation

The test item was tested for mutagenic potential using in vitro bacterial reverse mutation test. The test substance did not exert mutagenic activity both in the presence and the absence of metabolic activation.
Executive summary:

The test item was tested for the mutagenic potential using in vitro bacterial reverse mutation test (Ames test)(corresponding to the OECD No. 471). A preliminary range-finding assay was performed using four strains of Salmonella typhimurium (TA100, TA1535, TA98 and TA1537) and one strain of E. Coli (WPuvrA) up to a maximum dose of 5.0 mg/plate to determine the optimal non-toxic test dose. Precipitates were not observed in any strain either with or without metabolic activation. Cytotoxicity (i.e., reduction in the background lawn) was not observed in any strain with or without metabolic activation. Criteria for a negative response were met for all tester strains with and without metabolic activation.

The concentrations tested in the confirmatory assay were 100, 250, 500, 1000, 2500 and 5000 µg/plate using the preincubation method. Precipitates were not observed in any strain either with or without metabolic activation. Cytotoxicity (i.e., reduction in the background lawn) was not observed in any strain either with or without metabolic activation. Criteria for a negative response were met for all tester strains with and without metabolic activation.

The test result is used as a key value in the hazard assessment.