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Diss Factsheets

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:
no data
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Acceptable, well-documented study report which meets basic scientific principles.

Data source

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

Materials and methods

Principles of method if other than guideline:
The test was carried out in accordance with:
- Ames BN et al (1973). Carcinogens are mutagens: A simple test system combining liver homogenates for activation and bacteria for detection. Proc Nat Acad Sci USA 70: 2281-2285.
- Ames BN et al (1975). Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mut Res 31: 347-364.
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
4H-3,1-benzoxazine-2,4(1H)-dione
EC Number:
204-255-0
EC Name:
4H-3,1-benzoxazine-2,4(1H)-dione
Cas Number:
118-48-9
Molecular formula:
C8H5NO3
IUPAC Name:
4H-3,1-benzoxazine-2,4(1H)-dione
Details on test material:
- Name of test material (as cited in study report): Isatosäureanydrid (Isatoic anhydride)
- Analytical purity: no data
- Impurities (identity and concentrations): no data
- Storage condition of test material: 4°C
No further data

Method

Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Metabolic activation system:
rat liver S-9
Test concentrations with justification for top dose:
0, 20, 100, 500, 2500, 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: no data
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
for all strains; with S9 mix
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: N-methyl-N'-nitro-N-nitroso-guanidine
Remarks:
for strains TA 100 and TA 1535; without S9 mix
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylenediamine
Remarks:
for strain TA 98; without S9 mix
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 9-aminoacridine chloride monohydrate
Remarks:
for strain TA 1537; without S9 mix
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

NUMBER OF REPLICATIONS: 3 plates per dose

Evaluation criteria:
In general, a substance to be characterized as positive in the Ames test has to fulfil the following requirements:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results
Statistics:
no data

Results and discussion

Test results
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:
cytotoxicity
Remarks:
>= 2000 µg/plate
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

No increase in the number of revertants was observed. A bacteriotoxic effect was observed from 2000 µg/plate onwards, depending on the strain and test conditions. The test substance was completely soluble; no precipitation was noted.

Applicant's summary and conclusion

Executive summary:

Report Summary

The substance Isatosäureanhydrid was tested for mutagenicity in the Ames test.

Strains: TA 1535, TA 100, TA 1537, TA 98

Dose range: 20 µg - 5000 µg/plate

Test conditions: Standard plate test without and with metabolic activation (S-9 mix)

Solubility: Complete solubility of the test substance in DMSO

Toxicity: Depending on the strain, experiment and test conditions, a bacteriotoxic effect was observed from about 2000 µg/plate onward.

Mutagenicity: An increase in the number of his+ revertants could not be observed either without S-9 mix or after the addition of a metabolizing system.

Assessment: According to the results of the present study, the test substance Isatosäureanhydrid (i.e. isatoic anhydride) is not mutagenic in the Ames test under the experimental conditions chosen here.