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

Endpoint summary

Additional information

Gen mutation in bacteria

In a Standard Plate Test according to Ames et al. (1973, 1975) and following a standardized test protocol, the test substance (unknown purity) did not cause increased numbers of revertants in the S. typhimurium strains TA 1535, TA 100, TA 1537 and TA 98, neither with nor without metabolic activation (S-9 mix) in a dose range of 20 - 5000 µg/plate. The substance was completely soluble in DMSO and caused cytotoxicity from 2000 µg/plate onwards, depending on strain and test conditions. The controls were all valid. According to the results of the present study, the test substance is not mutagenic in the Ames test under the experimental conditions chosen here (BASF 1985).

In vivo

Cytogenicity in mammalians

A micronucleus test was performed following GLP requirements and OECD test guideline 474 in order to investigate the potential of the test substance to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse.

The test article was formulated in polyethylene glycol 400. This vehicle was used as negative control. The volume administered orally was 10 ml/kg bw 24 h and 48 h after a single application of the test article the bone marrow cells were collected for micronuclei analysis. 1000 polychromatic erythrocytes (PCE) per treated NMRI mouse (5 males, 5 females per test group) were evaluated for the occurrence of micronuclei.

At the 24 h preparation interval, dose levels of 200, 670 and 2000 mg/kg bw were investigated, and at the 48 h preparation interval only the dose level of 2000 mg/kg bw was investigated.

2 out of 24 treated animals of the high dose died. After treatment with the test article the number of NCEs was not increased as compared to the corresponding negative controls thus indicating that the test substance had no cytotoxic effectiveness. In comparison to the corresponding negative controls there was no significant enhancement in the frequency of the detected micronuclei at any preparation interval after application of the test article and with any dose level used. Suitable sensivity of the test system was given since a positive control substance (30 mg/kg bw cyclophosphamide) caused a distinct increase of induced micronucleus frequency.

In conclusion, it can be stated that during the study described and under the experimental conditions reported, the test article did not induce micronuclei as determined by the micronucleus test with bone marrow cells of the mouse. Therefore, the test substance is considered to be non-mutagenic in this micronucleus assay (CCR/BASF 1992).

Short description of key information:
in vitro
Gene mutation in bacteria
Ames test, S. typhimurium TA TA 1535, TA 1537, TA 98 and TA 100, with & without metabolic activation: negative (standardized protocol, according to Ames et al. 1973/1975; BASF 1985)

in vivo
Cytogenicity in mammalians
Micronucleus Test, NMRI mouse, oral: negative (GLP, OECD 474; CCR/BASF 1992)

Endpoint Conclusion:

Justification for classification or non-classification

Due to the negative results of an in vitro gene mutation test (Ames test) and an in vivo cytogenicity test (Micronucleus test), there is no indication given for a classification for the mutagenic potential of the test substance according to 67/548/EEC and GHS requirements.