Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference Type:
study report

Materials and methods

Test guideline
according to guideline
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
GLP compliance:
yes (incl. QA statement)
Type of assay:
bacterial forward mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
1,3,6,8-Pyrenetetrasulfonic acid, sodium salt
EC Number:
Cas Number:
Molecular formula:
1,3,6,8-Pyrenetetrasulfonic acid, sodium salt


Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Species / strain / cell type:
E. coli WP2 uvr A
Metabolic activation:
with and without
Metabolic activation system:
S9 mix (cofactor supplemented post-mitochondrial fraction)
Test concentrations with justification for top dose:
The test substance was formulated as a solution in sterile water (0.0158, 0.05, 0.158, 0.5, 1.58, 5, 15.8 and 50 mg/mL) to provide corresponding dose levels of up to 5000 µg/plate. The solutions were vortexed prior to use.
Vehicle / solvent:
The test substance was found to be soluble in sterile water that was used as the vehicle control.

Results and discussion

Test results
Key result
Species / strain:
other: TA1535, TA1537, TA98 , TA100 or E.coli WP2 uvrA
Metabolic activation:
with and without
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
Positive controls validity:
Remarks on result:
other: Based on these findings and on the evaluation system used, PTSA did not elicit evidence of bacterial mutagenicity in the Ames assay.

Applicant's summary and conclusion

Revertant colony counts for each strain are presented in Tables 1-5. Historical Control Data is presented in Appendix A.
The mean revertant colony counts for each strain treated with the vehicle were close to or within the expected range, considering the laboratory historical control range and/or published values (Mortelmans & Zeiger, 2000; Gatehouse, 2012). The positive control substances caused the expected substantial increases in revertant colony counts in both the absence and presence of S9 in each phase of the test confirming the sensitivity of the test and the activity of the S9 mix. Therefore, each phase of the test is considered valid.
No signs of contamination, toxicity or precipitation were noted in any of the strains. For all strains, at least eight non-toxic doses were evaluated; therefore bacterial mutagenicity was adequately assessed.
There was no concentration-related or substantial test substance related increases in the number of revertant colonies observed with strains TA1535, TA1537, TA98 , TA100 or E.coli WP2 uvrA in both the absence and presence of S9 using either the plate incorporation or the pre-incubation method.