Registration Dossier

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2018

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes (incl. certificate)
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Specific details on test material used for the study:
TEST MATERIAL NAME (as stated in study report): Intermediate F-37

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Valtris Specialty Chemicals / Batch/Lot No. W042272
- Expiration date of the lot/batch: November 21, 2019
- Purity: 84.5%

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Ambient temperature. In original container as supplied by the Sponsor. Containers were kept tightly closed in a dry, cool and well ventilated place.

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
Details on mammalian cell type (if applicable):
CELLS USED
- Source of cells: Molecular Toxicology Inc., 157 Industrial Park Dr. Boone, NC 28607, U.S.A.
Metabolic activation:
with and without
Metabolic activation system:
Arochlor 1254-induced rat liver S9 fraction
Test concentrations with justification for top dose:
Initial Toxicity-Mutation Test: 0.0015 - 5 µL/plate
Confirmatory Mutation Test: 0.13 - 4 µL/plate

The top dose in the confirmatory mutation test was chosen based on cytotoxicity observed in the initial toxicity-mutation test.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: solubility
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
mitomycin C
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium; in agar (plate incorporation)

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: three

DETERMINATION OF CYTOTOXICITY
- Method: inhibition of the background bacterial lawn and/or reduction in the number of revertant colonies
Rationale for test conditions:
per guidelines
Evaluation criteria:
per guidelines
Statistics:
Simple linear regression analysis was performed to assess the dose dependent nature of any increase in revertant colonies.

Results and discussion

Test results
Species / strain:
other: TA1537, TA1535, TA98, TA100, TA102
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:
Precipitation was not observed at the concentration of 5 µL/plate.

Applicant's summary and conclusion

Conclusions:
Under guideline (OECD 471) test conditions, Intermediate F37 was not mutagenic in the Bacterial Reverse Mutation Test at concentrations up to 5 µL/plate, with and without exogenous metabolic activation.