3-chloropropyne

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:

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GLP compliant guideline study, available as unpublished report, minor restriction in design (no E.coli/WP2 strain or S.typhimurium TA102 strain used) and/or reporting but otherwise adequate for assessment; however,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 guideline

Test guideline 1

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

Test guideline 2

Qualifier:

according to guideline

Guideline:

other: EU Method B.14 (Salmonella typhimurium - reverse mutation assay)

Test guideline 3

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

Test material information

Constituent 1

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.

Controls

Controls 1

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

Controls 2

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

Controls 3

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 results

Test results 1

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

Test results 2

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

Test results 3

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

Test results 4

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