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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1986
Report Date:
1986

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
no
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
liquid
Details on test material:
- Name of test material (as cited in study report): FMC 495
- Physical state: Viscous Colorless Liquid
- Analytical purity: > 98%
- Lot/batch No.: 1st Sample: MLS-2 ; 27, and 2nd Sample: MLS-B#86-15 T/T6306 MLS-4 ; 38
- Stability: Not completely defined
- Storage Conditions: Refrigerate
- Date Study Started: June 24, 1985
- Date Study Completed: May 1, 1986

Method

Target gene:
Not applicable
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Species / strain / cell type:
S. typhimurium TA 1538
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
1st Phase (preliminary test with TA100): 10.0, 33.3, 66.7, 100.0, 333.3, 666.7, 1000.0, 3333.3, 6666.7, and 10000.0 µg/plate with or without S-9 Activation.
2nd Phase with the 1st sample of DAP:
- mutagenicity assay with all five strains: 50, 100, 250, 500, and 1000 µg/plate with or without S9.
- repeat mutagenic assay: 250, 500, 1000, 1500, and 3000 µg/plate without S9.
3rd phase with the 2nd sample of DAP:
- mutagenicity assay: 150, 300, 600, 1500, 3000, and 6000 µg/plate without metabolic activation.
25, 50, 100, 250, 500, and 1000 µg/plate with metabolic activation.
Vehicle / solvent:
Dimethylsulphoxide (DMSO)
Controlsopen allclose all
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
5 µg/plate Migrated to IUCLID6: TA1535
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
75 µg/plate Migrated to IUCLID6: TA1537
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
5 µg/plate Migrated to IUCLID6: TA1538; TA98
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 2-Anthramine for all strings
Remarks:
4 µg/plate
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: ovenight at 37 ± 3 ˚C
- Exposure duration: 48 - 72 h at 37 ± 3 ˚C
- Expression time (cells in growth medium): not applicable
- Selection time (if incubation with a selection agent): not applicable
- Fixation time (start of exposure up to fixation or harvest of cells): not applicable

SELECTION AGENT (mutation assays): histidine

NUMBER OF REPLICATIONS: Triplicate

NUMBER OF CELLS EVALUATED: not applicable
Evaluation criteria:
For the test article to be considered positive, it must cause at least a doubling in the mean number of revertants per plate of at least one strain. This increase in the mean umber of revertants per plate must be accompanied by a dose response to increasing concentrations of the test article. For TA1537 or TA1538 revertants less than three-fold, the response must be reproducible.
Statistics:
For each triplicate plating, an average and standard deviation were calculated.

Results and discussion

Test results
Key result
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
The positive response of TA 1535 to DAP exposure was more than 2-fold and repeated in all tests performed for both samples.
Remarks on result:
other: strain/cell type: TA 1535

Applicant's summary and conclusion

Conclusions:
Interpretation of results: positive without metabolic activation For one out of five strains tested

Diallyl phthalate caused a positive response in S. typhimurium strain TA1535.