N,N'-bis[2-hydroxy-3-(2,2,3,3-tetrafluoropropoxy)propyl]-N,N,N',N'-tetramethylethane-1,2-diaminium dichloride

Genetic toxicity: in vitro

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was performed according to GLP and was designed to meet the requirements of OECD Guidelines for Testing of Chemicals No. 471 (adopted 1997, Bacterial Reverse Mutation Assay), and ICH Guidelines S2A and S2B (ICH 1995, 1997). No deviations or deficiencies that affected the integrity of the study are cited in the study report. The study is selected as the key study because the information provided for the hazard endpoint is sufficient for the purpose of classification and labelling and/or risk assessment.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-bis[2-hydroxy-3-(2,2,3,3- tetrafluoropropoxy)propyl]-N,N,N',N'-tetramethylethane-1,2- diaminium dichloride

IUPAC Name:

N,N'-bis[2-hydroxy-3-(2,2,3,3- tetrafluoropropoxy)propyl]-N,N,N',N'-tetramethylethane-1,2- diaminium dichloride

Constituent 2

Reference substance name:

1,1,2,2,17,17,18,18-octafluoro-6,13-dihydroxy-8,8,11,11-tetramethyl-4,15-dioxa-8,11-diazaoctadecane-8,11-diium dichloride

EC Number:

700-737-4

Cas Number:

1220100-43-5

Molecular formula:

C18H34F8N2O4.2Cl

IUPAC Name:

1,1,2,2,17,17,18,18-octafluoro-6,13-dihydroxy-8,8,11,11-tetramethyl-4,15-dioxa-8,11-diazaoctadecane-8,11-diium dichloride

Details on test material:

The test article was supplied by the Sponsor as a transparent, colorless, aqueous solution of test substance (white solid) at an analytically determined concentration of 47.24 mg/mL.

Method

Target gene:

Histidine (Salmonella) / Tryptophan (E. coli)

Species / strain

Species / strain 1

Species / strain / cell type:

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

Species / strain 2

Species / strain / cell type:

E. coli WP2 uvr A

Metabolic activation:

with and without

Metabolic activation system:

metabolic activation by liver extract (S-9) from rats treated with Arochlor 1254

Test concentrations with justification for top dose:

Initial mutagenicity assay at doses of 1.60, 5.00, 16.0, 50.0, 160, 500, 1600, and 5000 µg/plate, with and without S9.
An independent confirmatory assay subsequently was performed at doses of 50.0, 160, 500, 1600, and 5000 µg/plate, with and without S9.

Vehicle / solvent:

Deionized water

Controls

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Remarks:

TA98 - 2-nitrofluorene;-S9: TA100, TA1535 - sodium azide; TA1537 - ICR-191; WP2uvrA - 4-nitroquinoline-N-oxide. +S9: TA98 - benzo[a]pyrene; TA100, TA1535, TA1537 - 2-aminoanthracene; WP2uvrA - 2-aminoanthracene

Details on test system and experimental conditions:

Design
The test article was evaluated in the initial mutagenicity assay at doses of 1.60, 5.00, 16.0, 50.0, 160, 500, 1600, and 5000 µg/plate, with and without S9. Positive and vehicle controls were evaluated concurrently. All test and control articles were evaluated in duplicate plates. An independent confirmatory assay subsequently was performed at doses of 50.0, 160, 500, 1600, and 5000 µg/plate, with and without S9. Positive and vehicle controls were evaluated concurrently, and all test and control articles were evaluated in triplicate plates.

Frequency and Route of Administration
Tester strains were exposed to the test article via the plate incorporation methodology originally described by Ames et al. (1975) and Maron and Ames (1983). This methodology has been shown to detect a wide range of classes of chemical mutagens. In the plate incorporation methodology, the tester strain, test article, and S9 mix (where appropriate) are combined in molten top agar, which is then overlaid onto a minimal bottom agar plate. Following incubation, revertant colonies are counted.

Evaluation criteria:

A test article is considered to have produced a positive response if it induces a dose dependent increase in revertant frequency that is above 2.0-fold vehicle control values for tester strains TA98, TA100, and WP2uvrA, or above 3.0-fold vehicle control values for tester strains TA1535 and TA1537. In addition, any response should be reproducible.

A test article is considered to have produced a negative response if no dose-dependent, >2.0-fold or >3.0-fold increases are observed in tester strains TA98, TA100, and WP2uvrA, or TA1535 and TA1537, respectively.

Criteria for an Equivocal Response
Even after repeated trials, a test article may produce results that are neither clearly positive nor clearly negative (e.g., responses that do not meet the dose-dependency or fold increase requirements, but are reproducible). In those rare instances, the test article may be considered to have produced an equivocal response.

Statistics:

The average revertants/plate and the standard deviation were calculated.

Results and discussion

Test results

Test results 1

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

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

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 specified

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

These results indicate the test substance was negative in the Bacterial Reverse Mutation Assay with a Confirmatory Assay under the conditions, and according to the criteria, of the test protocol.

Executive summary:

The objective of this study was to evaluate the test article, and/or its metabolites, for their ability to induce reverse mutations at the histidine locus in several strains of Salmonella typhimurium (Salmonella; TA98, TA100, TA1535, and TA1537), and at the tryptophan locus of Escherichia coli (E. coli) strain WP2uvrA in the presence or absence of an exogenous mammalian metabolic activation system (S9).

The substance was evaluated in the initial mutagenicity assay, in all five tester strains, at doses of 1.60, 5.00, 16.0, 50.0, 160, 500, 1600, and 5000 µg/plate with and without S9. All doses of the test article as well as the concurrent positive and vehicle controls were evaluated in duplicate plates. Normal growth was observed in all five tester strains at all doses evaluated with and without S9. In addition, the test article was found to be freely soluble in the aqueous top agar at all doses evaluated with and without S9. Revertant frequencies for all doses of test substance, in all tester strains with and without S9, approximated or were less than those observed in the concurrent vehicle control cultures.

The substance was re-evaluated in the confirmatory mutagenicity assay at doses of 50.0, 160, 500, 1600, and 5000 µg/plate with and without S9. All doses of the test article, as well as the concurrent positive and vehicle controls, were evaluated in triplicate plates. Normal growth again was observed in all five tester strains at all doses evaluated with and without S9. In addition, the test article again was found to be freely soluble in the aqueous top agar at all doses evaluated with and without S9. Revertant frequencies for all doses of test substance, in all tester strains with and without S9, again approximated or were less than control values.

All positive and vehicle control values were within acceptable ranges, and all criteria for a valid study were met.

These results indicate the test substance was negative in the Bacterial Reverse Mutation Assay with a Confirmatory Assay under the conditions, and according to the criteria, of the test protocol.