N,N'-diacetylhydrazine

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:

weight of evidence

Study period:

not reported

Reliability:

3 (not reliable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study is a screening study which did not formally follow a guideline method, and was not formally performed under audited GLP, however, the laboratory facilities were operated according to GLP. The documentation in the report was very limited. Although the results are considered valid, the screening study itself cannot be considered formally reliable due to the lack of formal guideline, quality assurance, and very limited documentation.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2004

Report date:

2004

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Principles of method if other than guideline:

The guidance is not explicitly mentioned in the study report, however, it is mentioned that the experiments were conducted under the same test conditions and according to the same principles as studies performed under GLP using the standard plate-incorporation assay protocol.

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N'-diacetylhydrazine

EC Number:

221-576-1

EC Name:

N,N'-diacetylhydrazine

Cas Number:

3148-73-0

Molecular formula:

C4H8N2O2

IUPAC Name:

N'-acetylacetohydrazide

Test material form:

not specified

Method

Target gene:

Genes required for synthesis of histidine (S. typhimurium) or tryptophan (E. coli)

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Species / strain 2

Species / strain / cell type:

E. coli WP2

Additional strain / cell type characteristics:

other: pKM101

Species / strain 3

Species / strain / cell type:

E. coli WP2 uvr A pKM 101

Metabolic activation:

with and without

Metabolic activation system:

S9-mix

Test concentrations with justification for top dose:

100 to 5000 µg/plate

Vehicle / solvent:

dried dimethyl sulphoxide

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

not specified

True negative controls:

not specified

Positive controls:

not specified

Positive control substance:

not specified

Details on test system and experimental conditions:

METHOD OF APPLICATION: in medium; in agar (plate incorporation); preincubation

First standard plate-incorporation assay with all strains with and without S9-mix
- Preincubation period: none
- Exposure duration: 3 days

NUMBER OF REPLICATIONS: not reported

NUMBER OF CELLS EVALUATED: not reported

DETERMINATION OF CYTOTOXICITY
- no signs of cytotoxicity are reported

A second study following the pre-incubation assay protocol in strains TA98 and TA100 with S9-mix
- Preincubation period: not reported
- Exposure duration: 3 days
- Expression time (cells in growth medium): 3 days

NUMBER OF REPLICATIONS: not reported

NUMBER OF CELLS EVALUATED: not reported

DETERMINATION OF CYTOTOXICITY
- no signs of cytotoxicity are reported

Evaluation criteria:

Increase in the number of revertant colonies per plate in at least one strain with or without with S9-mix

Statistics:

not reported

Results and discussion

Test results

Species / strain:

other: TA1535, TA 1537, TA98, TA100, WP2(pKM101), WP2 uvrA (pKM101)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

not specified

Untreated negative controls validity:

not specified

Positive controls validity:

not specified

Additional information on results:

no additional information on results reported

Remarks on result:

other: all strains/cell types tested

Applicant's summary and conclusion

Conclusions:

Interpretation of results:
negative

The test substance was non-mutagenic in a bacterial reverse mutation assay in both the presence and absence of metabolic activation (S9-mix). Although the result is considered to be valid, due to the lack of formal guideline and quality assurance, and very limited documentation, it should be regarded as not being reliable.

Executive summary:

The mutagenic potential of the test substance was studied in a non-GLP bacterial reverse mutation assay for Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strains WP2 (pKM101) and WP2 uvrA (pKM101) by applying the standard plate-incorporation assay protocol in accordance with OECD Testing Guideline No. 471. The test was conducted in the absence and presence of the metabolic activation system S9-mix derived from rat liver. In addition, a second experiment using the pre-incubation assay protocol in accordance with OECD Testing Guideline No. 471 was conducted with S. typhimurium strains TA98 and TA100 in the presence of S9-mix. The substance was tested in the concentration range from 100 to 5000 µg/plate and plates were assessed after incubation for 3 days. All tests gave negative, i.e. non-mutagenic, response in both the presence and absence of S9-mix.