(1E)-1-chloro-3,3,3-trifluoroprop-1-ene

Genetic toxicity: in vitro

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:

2011

Report date:

2011

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Test guideline 2

Qualifier:

according to guideline

Guideline:

JAPAN: Guidelines for Screening Mutagenicity Testing Of Chemicals

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Test material form:

gas under pressure: liquefied gas

Details on test material:

- Storage Conditions: Room temperature, upright in the dark without desiccant

Method

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 uvr A

Metabolic activation:

with and without

Metabolic activation system:

Aroclor 1254-induced rat liver S9

Test concentrations with justification for top dose:

- Preliminary toxicity assay: 1, 3, 6, 8, 11, 21 and 37 mmoles/L (24,500; 73,400; 147,000; 196,000, 269,000; 513,000 and 905,000 ppm, respectively)
- Mutagenicity and confirmatory assay: 3, 6, 8, 11, 21 and 37 mmoles/L (73,400; 147,000; 196,000, 269,000; 513,000 and 905,000 ppm, respectively).
- Confirmatory retest: 0.7, 1, 3, 6, 8, 11, 21 and 37 mmoles/L (17,200; 24,500; 73,400; 147,000; 196,000, 269,000; 513,000 and 905,000 ppm, respectively)

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

no

True negative controls:

no

Positive controls:

yes

Positive control substance:

9-aminoacridine

2-nitrofluorene

sodium azide

methylmethanesulfonate

other: 2-aminoanthracene

Details on test system and experimental conditions:

METHOD OF APPLICATION: test system was exposed to the test article via the desiccator methodology, a modification of the plate incorporation methodology originally described by Ames et al. (1975) and updated by Maron and Ames (1983). This test system has been shown to detect a wide range of classes of chemical mutagens (McCann et al., 1975; McCann and Ames, 1976). The desiccator methodology has been shown to be an effective method for detecting the genotoxic activity of volatile and gaseous test articles (Wagner et al., 1992). Briefly, once agar had solidified, the plates were inverted in 9 liter dessicators and an appropriate quantity of test material was introduced into each dessicator by withdrawing the appropriate amount of air and replacing with test material.

DURATION
- Original assay: following 24 hour exposure to test article, the plates were removed from the dessicators and allowed to inclubate at 37 °C for an additional 24 hours.
- Confirmatory assay: exposure duration 48 hours in dessicator.

NUMBER OF REPLICATIONS: triplicate

DETERMINATION OF CYTOTOXICITY
- Method: relative total growth; number of revertants

Evaluation criteria:

- Data sets for tester strains TA1535 and TA1537 were judged positive if the increase in mean revertants at the peak of the dose response was equal to or greater than 3.0-times the mean vehicle control value.
- Data sets for tester strains TA98, TA100 and WP2 uvrA were judged positive if the increase in mean revertants at the peak of the dose response was equal to or greater than 2.0-times the mean vehicle control value.
- An equivocal response is a biologically relevant increase in a revertant count that partially meets the criteria for evaluation as positive. This could be a dose-responsive increase that does not achieve the respective threshold cited above or a non-dose responsive increase that is equal to or greater than the respective threshold cited.
- A response will be evaluated as negative, if it is neither positive nor equivocal.

Results and discussion

Test results

Test results 1

Key result

Species / strain:

other: Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

True negative controls validity:

not applicable

Positive controls validity:

valid

Test results 2

Key result

Species / strain:

E. coli WP2 uvr A

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

cytotoxicity

Vehicle controls validity:

not applicable

Untreated negative controls validity:

valid

True negative controls validity:

not applicable

Positive controls validity:

valid

Additional information on results:

PRELIMINARY TOXICITY ASSAY
- No background lawn toxicity was observed however a reduction in revertant counts was observed beginning at 11, 21 or at 37 mmoles/L with some test conditions.

MUTAGENICITY ASSAY: No positive mutagenic responses were observed with any of the tester strains in the presence or absence of S9 activation. No precipitate was observed. Toxicity was observed beginning at 11, 21 or at 37 mmoles/L.

CONFIRMATORY MUTAGENICITY ASSAY
No positive mutagenic response was observed with tester strains TA100, TA1535 and WP2 uvrA in the absence of S9 activation and with tester strains TA100, TA1535 and TA1537 in the presence of S9 activation. No precipitate was observed. Toxicity was observed beginning at 21 or 37 mmoles/L. Tester strain TA98 in the presence and absence of S9 was not evaluated due to confluent growth but was retested. Due to excessive toxicity, as a result of a drop in the revertant counts beginning at 8 mmoles/L, tester strain TA 1537 in the absence of S9 was retested. Due to an unacceptable vehicle control value, tester strain WP2 uvrA in the absence of S9 activation was not evaluated but was retested. In the retest of the confirmatory mutagenicity assay, no positive responses were observed with tester strains TA98 and TA1537 in the absence of S9 activation and with tester strains TA98 and WP2 uvrA in the presence of S9 activation. No precipitate was observed. Toxicity was observed beginning at 21 or 37 mmoles/L.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

		-S9		+S9
Strain	Test conc (mmol/L)	Average reverants original	Average revertants – confirmatory	Average reverants – original	Average revertants – confirmatory
TA98	Vehicle	35	30	24	12
	0.7	NA	31	NA	9
	1	NA	34	NA	9
	3	27	29	15	10
	6	28	24	25	9
	8	22	27	18	11
	11	19	22	13	10
	21	4	15	3	10
	37	0	0	0	0
	Positive control	129	351	442	408
TA100	Vehicle	179	96	209	106
	3	214	108	216	88
	6	179	113	203	83
	8	184	130	236	72
	11	216	88	214	63
	21	205	40	267	40
	37	0	0	0	0
	Positive control		772		407
TA1535	Vehicle	10	11	7	9
	3	10	8	7	10
	6	9	9	6	5
	8	11	11	10	12
	11	8	7	8	7
	21	6	11	5	7
	37	0	0	0	0
	Positive control	293	666	73	140
TA1537	Vehicle	7	10	7	7
	0.7	NA	6	NA	NA
	1	NA	6	NA	NA
	3	3	10	4	6
	6	3	11	3	6
	8	5	7	4	4
	11	0	6	7	5
	21	1	4	0	5
	37	0	0	0	0
	Positive control	992	633	36	473
WP2 uvrA	Vehicle	14	45	19	42
	0.7	NA	NA	NA	39
	1.0	NA	NA	NA	42
	3	13	56	23	43
	6	12	55	16	28
	8	11	39	13	35
	11	4	33	6	32
	21	2	15	5	34
	37	0	8	0	4
	Positive control	168	372	181	198

Applicant's summary and conclusion

Conclusions:

The results of the bacterial reverse mutation test using gas-phase exposure indicate that, under the conditions of this study, the test article did not cause a positive mutagenic response in either the presence or absence of Aroclor induced rat liver S9.

Executive summary:

In a reverse mutation assay, performed according to OECD Guideline 471 and GLP, Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100 and Escherichia coli strain WP2uvrA were treated with the test material using the desiccator methodology, a modification of the Ames plate incorporation methodology, in the presence and absence of S9 -mix. The assay was performed in three phases: a preliminary toxicity assay, a mutagenicity assay and a confirmatory mutagenicity assay. Based on the findings of the preliminary toxicity assay, the maximum dose plated in the confirmatory mutagenicity assay was 37 mmoles/L (905,000 ppm). In the mutagenicity and confirmation assay, no precipitation was observed and toxicity was observed beginning at 11, 21 or at 37 mmoles/L. In the mutagenicity and confirmation assay, no positive mutagenic response was observed. In conclusion, under the conditions of this study, the test article did not cause a positive mutagenic response in either the presence or absence of Aroclor-induced rat liver S9.