4H-3,1-benzoxazine-2,4(1H)-dione

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

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

Test material information

Constituent 1

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

Controls

Controls 1

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

Controls 2

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

Controls 3

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

Controls 4

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.