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Toxicological information

Genetic toxicity: in vitro

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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
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
GLP compliant guideline study, available as unpublished report, minor restriction in design (no E.coli/WP2 strain or S.typhimurium TA102 strain used) and/or reporting but otherwise adequate for assessment; however, no details on purity of the test material are given

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1990
Report date:
1990

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Deviations:
yes
Remarks:
no E. coli/WP2 strain or S. typhimurium TA102 strain used
Qualifier:
according to guideline
Guideline:
other: EU Method B.14 (Salmonella typhimurium - reverse mutation assay)
Qualifier:
according to guideline
Guideline:
other: Environmental Protection Agency, Code of Federal Regulations, Title 40, Subpart F-Genetic Toxicity, Revision July 1, 1986 “The Salmonella Typhimurium Reverse Mutation Assay"
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
3-chloropropyne
EC Number:
210-856-9
EC Name:
3-chloropropyne
Cas Number:
624-65-7
Molecular formula:
C3H3Cl
IUPAC Name:
3-chloroprop-1-yne
Details on test material:
- Name of the test material (as cited in study report): Propargylchloride
- Batch No.: P8/87 (Lab. - No.54/L)
- Physical state: yellow liquid
- Stability, pure: 6 months
- Stability, in solvent: in water, ethanol, methanol, acetone, DMSO, DMF and Toluene >24 hours
- Storage: room temperature
- Expiration date: 1989-12-01

Method

Target gene:
His-locus
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 1535, TA 1537, TA 1538, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Liver S9 fraction of Aroclor - induced rats
Test concentrations with justification for top dose:
- without S9 mix: 10.0, 33.3, 100.0, 333.3, 666.6, 1000.0 µg/plate
- with S9 mix: 33.3, 100.0, 333.3, 666.6, 1000.0, 5000.0 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Toluene
- Justification for choice of solvent/vehicle: It was chosen to its solubility properties and its relative nontoxicity for the bacteria.
Controlsopen allclose all
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
Toluene
True negative controls:
no
Positive controls:
yes
Remarks:
10 µg/plate
Positive control substance:
sodium azide
Remarks:
TA1535, TA100; without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
Toluene
True negative controls:
no
Positive controls:
yes
Remarks:
50 µg/plate
Positive control substance:
other: 4-nitro-o-phenylene-diamine
Remarks:
TA1537, TA1538, TA98; without metabolic activation
Untreated negative controls:
yes
Negative solvent / vehicle controls:
yes
Remarks:
Toluene
True negative controls:
no
Positive controls:
yes
Remarks:
10 µg/plate
Positive control substance:
other: 2-aminoanthracene
Remarks:
TA1535, TA1537, TA1538, TA98, TA100; with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Exposure duration: 72 h

NUMBER OF REPLICATIONS:
- Experiment performed in duplicate
- Three plates per condition per experiment

DETERMINATION OF CYTOTOXICITY:
- Method: other: growth inhibition of revertant clones

DATA RECORDING
- The colonies were counted using the BIOTRAN III counter
Evaluation criteria:
- A test article is considered as positive if either a significant dose-related increase in the number of revertants or a significant and reproducible increase for at least one test concentration is induced.
- A test article producing neither a significant dose-related increase in the number of revertants nor a significant and reproducible positive response at any one of the test points is considered non-mutagenic in this system.
- A test article is considered as mutagen if in strain TA 100 the number of reversions is at least twice as high and in strains TA 1535, TA 1537, TA 1538, and TA 98 it is at least three times higher as compared to the spontaneous reversion rate.
- Also, a dose-dependent increase in the number of revertants is regarded as an indication of possibly existing mutagenic potential of the test article regardless whether the highest dose induced the above described enhancement factors or not.
Statistics:
No appropriate statistical method was available.

Results and discussion

Test resultsopen allclose all
Species / strain:
S. typhimurium, other: TA1535, TA100
Metabolic activation:
with and without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
without
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Species / strain:
S. typhimurium, other: TA1537, TA1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:
RANGE-FINDING/SCREENING STUDIES:
To evaluate the toxicity of the test article a pre-study was performed with strains TA 98 and TA 100.
The plates with the test article showed normal background growth up to 5000.0 µg/plate in strain TA 98 and TA 100, respectively.
However, no spontaneous revertants occurred at 5000.0 µg/plate in strain TA 98 and at 1000.0 and 5000.0 µg/plate in strain TA 100, all without metabolic activation.

ADDITIONAL INFORMATION ON CYTOTOXICITY:
Toxic effects, evidenced by a reduction in the number of spontaneous revertants, occurred in strain TA 1537 at 100.0 and 333.3 µg/plate without S9 mix and at 5000.0 µg/plate with S9 mix (exp. II). Also in strain TA 1538 at 5000.0 µg/plate with S9 mix (exp. II).
The plates incubated with the test article showed normal background growth up to 1000.0 µg/plate without S9 mix and up to 5000.0 µg/plate in the presence of S9 mix in all strains used.

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
positive