Registration Dossier

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: Acceptable, well documented study report, GLP conform

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1985
Report Date:
1985

Materials and methods

Test guideline
Qualifier:
no guideline followed
Principles of method if other than guideline:
Ames B.N. et al.: Proc. Nat. Acad. Sci. USA 70, 2281 - 2285, 1973
Ames B.N. et al.: Mutation Res. 31, 347-364, 1975
Green, M.H.L. and Muriel, W.J.: Mutation Research 38, 3-32, 1976
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Naphthalimid
- Physical state: grey powder
- Analytical purity: 96.1%
- Impurities (identity and concentrations): Trimellitacid, Hemimellitacid as Ammonium salt (Amide, Imide)
- Storage condition of test material: dark at 22°C
- pH: 5.8

Method

Target gene:
S. typhimurium: his-
E. coli: trp-
Species / strain
Species / strain:
other: S. typhimurium TA 100, TA 1535, TA 1537, TA 1538, TA 98; Escherichia coli WP2uvrA
Additional strain characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
S9 mix from Aroclor 1254 induced rat livers
Test concentrations with justification for top dose:
0.8 to 10000 µg/plate, differing between strains; per strain and experiment 5 to 6 doses (in total 9 different doses)
Vehicle:
DMSO
Controls
Negative controls:
no
Solvent controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
2-nitrofluorene
sodium azide
benzo(a)pyrene
other: N-Methyl-N´-nitro-N-nitrosoguanidine (MNNG), 2-Aminoanthracene
Remarks:
see "details on test system"
Details on test system and conditions:
METHOD OF APPLICATION: in agar (plate incorporation), 2 independent experiments

DURATION
- Preincubation period: overnight
- Exposure duration: 48 - 72 h at 37°C

NUMBER OF REPLICATIONS: 2

POSITIVE CONTROL SUBSTANCES:
without metabolic activation:
Na-azide: TA 100, TA 1535
9-Aminoacridine: TA 1537
2-Nitrofluorene: TA 98, TA 1538
N-Methyl-N´-nitro-N-nitrosoguanidine (MNNG): WP2uvrA

with metabolic activation:
Benzo[a]pyrene: TA 98, TA 100, TA 1535, TA 1537, TA 1538, WP2uvrA
2-Aminoanthracene: TA 98, TA 100, TA 1535, TA 1537, TA 1538, WP2uvrA

Results and discussion

Test resultsopen allclose all
Species / strain:
other: S. typhimurium TA 100, TA 1535, TA 1537, TA 1538, TA 98; Escherichia coli WP2uvrA
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity:
yes
Remarks:
The test substance proved to be toxic >= 500 ug/plate
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Remarks on result:
other: strain/cell type: S. typhimurium TA 100; E. coli WP2uvrA
Remarks:
Migrated from field 'Test system'.
Species / strain:
other: S. typhimurium TA 100, TA 1535, TA 1537, TA 1538, TA 98; Escherichia coli WP2uvrA
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
The test substance proved to be toxic >= 500 ug/plate
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Remarks on result:
other: strain/cell type: S. typhimurium TA 1535, TA 1537, TA 1538, TA 98
Remarks:
Migrated from field 'Test system'.
Species / strain:
other: S. typhimurium TA 100, TA 1535, TA 1537, TA 1538, TA 98; Escherichia coli WP2uvrA
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity:
yes
Remarks:
The test substance proved to be toxic >= 500 ug/plate
Vehicle controls valid:
yes
Negative controls valid:
not examined
Positive controls valid:
yes
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Mutagenicity tests

Experiment 1
Strain (dose range in µg/plate) Metabolic activation system mean revertants in Controls maximum revertant factor* dose dependency Assessment
TA 98 no 38 0.8 no negative
(4-10,000) yes 41 1.3 no negative
TA 100 no 198 0.8 no negative
(4-10,000) yes 174 2.0 ambiguous** positive
TA 1535 no 18 1.1 no negative
(4-10,000) yes 13 1.5 no negative
TA 1537 no 12 1.3 no negative
(4-10,000) yes 9 1.6 no negative
TA 1538 no 25 1.5 yes negative
(4-10,000) yes 22 1.0 no negative
WP2 uvr A no 16 1.1 no negative
(4-10,000) yes 16 3.6 yes positive
Experiment 2
Strain (dose range in µg/plate) Metabolic activation system mean revertants in Controls maximum revertant factor* dose dependency Assessment
TA 98 no 25 1.0 no negative
(4-2,500) yes 27 1.3 no negative
TA 100 no 149 1.0 no negative
(4-2,500) yes 151 2.1 ambiguous** positive
TA 1535 no 22 0.7 no negative
(4-2,500) yes 13 1.1 no negative
TA 1537 no 14 1.1 no negative
(4-2,500) yes 15 0.9 no negative
TA 1538 no 11 1.4 no negative
(4-2,500) yes 18 1.1 no negative
WP2 uvr A no 18 1.1 no negative
(4 -2,500) yes 17 2.5 ambiguous** positive
* only in non-precipitating, non-toxic doses

** in all cases, the dose-dependency showed the following scheme: dose-dependent increase of revertants up to second highest dose without precipitation accompanied by slight to very slight decrease of the maximum revertant number in the highest non-precipitating dose

Applicant's summary and conclusion

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