Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

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:
1985-04-03 to 1985-12-02
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Conduction and documentation of study very acceptable. Study report available.

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1985
Report Date:
1985
Reference Type:
publication
Title:
Unnamed
Year:
2000
Report Date:
2000

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
Principles of method if other than guideline:
Test performed according to standard protocols by inclusion of a metabolic activation system (S9-Mix from livers of Sprague-Dawley rats pretreated with Aroclor 1254).
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): 2,4-Pentanedione
- Physical state: liquid
- Expiration date of the lot/batch: not reported
- Stability under test conditions: stable

Method

Target gene:
not applicable
Species / strain
Species / strain / cell type:
other: S. typhimurium, TA 98, 100, 1535, 1537, 1538
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with
Metabolic activation system:
S9-Mix
Test concentrations with justification for top dose:
0.3 - 30mg/plate
Vehicle / solvent:
water
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4 -nitro-o-phenylenediamine
Remarks:
TA98 and TA1538 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
TA100 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
TA1537 without S9
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
all strains with S9
Details on test system and experimental conditions:
METHOD OF APPLICATION:in agar (plate incorporation)


DURATION
- Exposure duration: 24 and 48 h



NUMBER OF REPLICATIONS:3



DETERMINATION OF CYTOTOXICITY
- Method: growth inhibition


Evaluation criteria:
Test chemicals which produce at least a 2-fold and dose-related increase in mutant colonies over the concurrent solvent control value are considered to be bacterial mutagens and suspect mammalian mutagens.
Statistics:
not reported

Results and discussion

Test results
Species / strain:
other: S. typhimurium, TA 98, 100, 1535, 1537, 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
97.8 mg/plate produced significant cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not applicable
Positive controls validity:
valid
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: none reported
- Effects of osmolality: none reported
- Evaporation from medium: none reported
- Water solubility: soluble
- Precipitation: none reported
- Other confounding effects: none reported


RANGE-FINDING/SCREENING STUDIES: yes, outside GLP


COMPARISON WITH HISTORICAL CONTROL DATA: yes, compliant
Remarks on result:
other: all strains/cell types tested
Remarks:
Migrated from field 'Test system'. Remarks: S. typhimurium, TA 98, 100, 1535, 1537, 1538

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

From the results it is concluded that 2,4-pentanedione is not mutagenic under the conditions of the assay.
Executive summary:

2,4-pentanedione (99.2% pure) was tested for potential mutagenic activity using the Salmonella/microsome bacterial mutagenicity assay (Ames test).

The test was performed according to standard protocols by inclusion of a metabolic activation system (S9-Mix from livers of Sprague-Dawley rats pretreated with Aroclor 1254). In preliminary trials with strain TA 100 only, a concentration of 97.4 mg/plate proved to completely inhibit bacterial growth. The next lower concentration of 30 mg/plate showed some toxic effects. Accordingly, doses selected on the basis of these trials were 0.3, 1.0, 3.0, 10.0 and 30.0 mg/plate. Incubations were run in triplicate at 37°C for 24 to 48 hours. Plates were examined for the condition of their background lawns and growth was recorded as either confluent (non-toxic), sparse (moderately toxic) or absent (toxic). The solvents of choice was water. Concurrent solvent and positive controls were run in each test. Positive control substances were without S9-mix 0.01 mg 4 -nitro-ophenylenediamine/ plate for TA98 and TA1538, 0.01 mg sodium azide for TA100, 0.06 mg 9-aminoacrididine/plate for TA1537 and with S9-mix 0.01 mg 2-aminoanthracene for all strains.

2,4-pentanedione did not produce a doubling or a dose-response relationship of the number of revertants/plate in the Salmonella typhimurium strains used neither in the absence nor in the presence of a metabolic activation system. Dose selection appeared to be in a suitable range, because in tests both with and without metabolic activation, bacteriotoxicty was observe d at 30 mg/plate (highest dose tested) with all strains. Well proven positive controls did produce mutagenic effects demonstrating the functionality of the test system. It can therefore be concluded that 2,4-pentanedione is not mutagenic under the conditions of the assay.