Trifluoroacetic anhydride

Genetic toxicity: in vitro

Administrative data

Endpoint:

in vitro gene mutation study in bacteria

Remarks:

Type of genotoxicity: gene mutation

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

1986-03-04 to 1986-03-25

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

The study was performed similarly to the OECD (No. 471) with some restrictions (only the 2-Anthramine is used as a positive control for the efficacy of S9 mix. Strain S. typhimurium TA102 or E.coli WP2 were not used. No data on the composition of the substance tested) and was in compliance with the GLP. Sodium Trifluoroacetate was tested instead of trifluoroacetic anhydride (TFAH) according to a reliable analogue approach in order to be free of the cytotoxic effect on bacteria due to the extreme acid pH of resulting Trifluoroacetic acid (TFA) (0.45) formed when TFAH is mixed to the aqueous culture medium.

Cross-reference

Cross-reference 1

Reason / purpose for cross-reference:

reference to same study

Cross-reference 2

Reason / purpose for cross-reference:

reference to other study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1986

Report date:

1986

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

: only the 2-Anthramine is used as a positive control for the efficacy of S9 mix. Strain S. typhimurium TA102 or E.coli WP2 were not used. No data on the composition of the substance tested.

Principles of method if other than guideline:

Not applicable

GLP compliance:

yes

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Sodium trifluoroacetate

EC Number:

220-879-6

EC Name:

Sodium trifluoroacetate

Cas Number:

2923-18-4

IUPAC Name:

sodium trifluoroacetate

Details on test material:

- Name of test material (as cited in study report): Sodium Trifluoroacetate (Trifluoroacetate de sodium, TFAS)
- Physical state: white powder
- Storage condition of test material: no data
No other information

Method

Target gene:

Each strains derived from S. thyphimurium LT2 contains one mutation in the histidine operon, resulting in a requirement for histidine. See details in Table 7.6.1/1.

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA1535, TA1537, TA1538, TA98 and TA100

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

other: additional mutation in uvrB and rfa genes. The plasmid pKM101 was present in TA98 and TA100 in order to increase the sensibility of these strains to some mutagens. See details in Table 7.6.1/1.

Metabolic activation:

with and without

Metabolic activation system:

S9 fraction was obtained from the liver of Sprague-Dawley male rats treated by the intraperitoneal route with Aroclor 1254 (500 mg/kg) dissolved in maize oil.

Test concentrations with justification for top dose:

Preliminary test: 0.01, 0.05, 0.1, 0.5, 1, 5, and 10 mg/plate.
Mutagenicity experiments: 0.1, 0.5, 1, 5 and 10 mg/plate.

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: no data

The susbtance is dissolved in DMSO at the highest concentration of 100 mg/mL. The succesive dilutions were performed from this initial solution.

Controls

Untreated negative controls:

yes

Remarks:

sterility controls for the substance's solution, the solvent and the S9 mix.

Negative solvent / vehicle controls:

yes

Remarks:

DMSO

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: 9-Aminoacridine (TA1537), 2-Nitrofluorene (TA98 and TA1538), Methyl methanesulphonate (TA 100) and Ethyl methanesulphonate (TA1535) for without S9 mix efficacy control. 2-Anthramine for with S9 mix efficacy control.See details in Table 7.6.1/3.

Details on test system and experimental conditions:

METHOD OF APPLICATION:
direct incorporation method: 100 µL of the DMSO diluted substance were incorporated in glass tubes. Then were added the bacteria (0.1 mL), the S9 fraction (0.5 mL) in the case of test with metabolic activation and the agar. After agitation the mix was plated on a Petri plate containing Vogel and Bonner medium.

DURATION
- Preincubation period: no data
- Exposure duration: 48 hours
SELECTION AGENT (mutation assays): not applicable
SPINDLE INHIBITOR (cytogenetic assays): not applicable
STAIN (for cytogenetic assays): not applicable
NUMBER OF REPLICATIONS: 3 plates/dose/strain. Two independent experiments were performed.
NUMBER OF CELLS EVALUATED: no data
DETERMINATION OF CYTOTOXICITY
- Method: no data
OTHER EXAMINATIONS:
- Determination of polyploidy: not required
- Determination of endoreplication: not required
- Other:
OTHER:

Evaluation criteria:

A reproducible 2-fold increase in the number of revertants compared with the vehicle controls was considered as a positive result. Reference to historical data, or other considerations of biological relevance may also be taken into account in the evaluation of the data obtained.

Statistics:

no data

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA1535, TA1537, TA1538, TA98 and TA100

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS: not applicable

RANGE-FINDING/SCREENING STUDIES: a preliminary study was performed on the TA100 strain without metabolic activation. The tested concentrations of the substance were 0.01, 0.05, 0.1, 0.5, 1, 5 and 10 mg/plate. No cytotoxicity in terms of decrease of the number of revertants or effect on the bacterial lawn was observed.

COMPARISON WITH HISTORICAL CONTROL DATA: The number of revertants for the vehicle and positive controls was similar to the historical data.
ADDITIONAL INFORMATION ON CYTOTOXICITY: no additional information

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Table 7.6.1/4:Number of revertants per plate (mean of triplicates) in the absence of metabolic activation in first test (direct plate incorporation method)

TFAS Concentration (mg/plate)	TA 1535		TA1537		TA 98		TA 100		TA 1538
	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation
0*	12.7	5.5	10.7	2.9	28.0	3.6	86.0	7.8	18.3	5.7
0.1	12.7	4.9	16.0	2.6	20.3	4.0	84.0	7.5	20.0	7.0
0.5	14.3	7.6	10.7	4.1	17.0	1.7	83.0	3.6	15.3	1.1
1	15.3	3.5	10.3	1.1	17.7	5.5	86.0	6.9	17.0	2.6
5	14.0	2.6	11.7	4.0	16.3	1.5	100.3	10.7	23.0	10.5
10	18.7	4.0	15.3	4.1	25.7	2.3	102.3	4.9	17.0	1.7
Positive control**	5723.0	-	989.0	-	264.5	-	301.0	-	199.0	-

*Solvent control = negative control: 100 µL DMSO

**Mutagens positive controls:

- 2.N.F. (0.001 mg/plate) in TA98 and TA1538 strains

- M.M.S. (0.1 mg/plate) in TA100 strain

- E.M.S. (10 mg/plate) in TA1535 strain

- 9 A.A. ( 0.05 mg/plate) in TA 1537 strain

 

 

 

 

 

 

 

Table 7.6.1/5:Number of revertants per plate (mean of triplicates) in the presence of metabolic activation (S9 mix) in first test (direct plate incorporation method)

TFAS Concentration (mg/plate)	TA 1535		TA1537		TA 98		TA 100		TA 1538
	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation
0*	15.7	8.1	12.0	2.6	27.3	8.5	107.7	13.0	32.0	2.6
0.1	20.3	2.3	11.3	1.1	29.0	5.0	106.3	16.0	36.0	7.0
0.5	19.3	1.5	19.3	6.6	28.0	1.0	99.3	3.0	27.7	4.1
1	15.3	4.0	12.3	2.9	31.3	2.5	94.7	6.6	30.0	9.1
5	15.0	3.0	15.0	3.0	31.7	5.8	98.7	12.3	28.0	3.0
10	17.3	3.2	11.3	0.6	26.7	6.5	109.3	13.0	26.7	11.0
Positive control**	225.0	-	150.5	-	1003.0	-	1453.0	-	831.0	-

*Solvent control = negative control: 100 µL DMSO

**Mutagens positive controls:

- 2.A. (0.002 mg/plate)

 

 

 Table 7.6.1/6:Number of revertants per plate (mean of triplicates) in the absence of metabolic activation in second test (direct plate incorporation method)

TFAS Concentration (mg/plate)	TA 1535		TA1537		TA 98		TA 100		TA 1538
	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation
0*	9.7	4.5	12.0	2.0	33.7	3.0	99.7	19.5	20.0	4.0
0.1	4.3	2.1	8.3	3.0	29.7	4.1	103.0	8.6	20.3	5.8
0.5	6.0	1.0	6.0	1.0	27.7	2.3	109.3	3.2	18.7	1.1
1	9.0	3.0	10.3	1.1	25.0	6.0	108.7	18.4	19.3	7.5
5	8.7	5.8	7.3	2.5	25.7	8.5	104.7	2.5	22.0	1.7
10	7.3	4.5	8.3	5.8	34.3	3.5	100.7	17.4	18.0	3.6
Positive control**	7301.5	-	445.0	-	263.0	-	450.5	-	254.5	-

*Solvent control = negative control: 100 µL DMSO

**Mutagens positive controls:

- 2.N.F. (0.001 mg/plate) in TA98 and TA1538 strains

- M.M.S. (0.1 mg/plate) in TA100 strain

- E.M.S. (10 mg/plate) in TA1535 strain

- 9 A.A. ( 0.05 mg/plate) in TA 1537 strain

 

Table 7.6.1/7:Number of revertants per plate (mean of triplicates) in the presence of metabolic activation (S9 mix) in second test (pre-incubation method)

 

TFAS Concentration (mg/plate)	TA 1535		TA1537		TA 98		TA 100		TA 1538
	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation	Mean	Standard deviation
0*	7.7	2.0	10.3	3.5	44.0	3.6	117.0	8.6	29.3	5.7
0.1	9.7	3.2	8.0	2.6	38.0	6.5	136.7	1.1	29.0	4.6
0.5	10.0	4.0	8.7	2.9	43.0	9.8	125.3	13.0	29.7	5.7
1	11.3	1.1	9.7	2.5	41.3	2.9	125.7	11.0	25.3	1.5
5	10.0	2.6	8.3	3.2	44.3	5.8	120.0	9.8	22.3	3.0
10	7.7	3.0	7.7	1.1	46.3	5.1	111.3	17.4	22.0	4.6
Positive control**	151.5	-	188.0	-	1510.0	-	2241.5	-	601.0	-

*Solvent control = negative control: 100 µL DMSO

**Mutagens positive controls:

- 2.A. (0.002 mg/plate)

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative with and without metabolic activation

Under the test conditions, the test item Sodium Trifluoroacetate did not show mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium.

Executive summary:

In a reverse gene mutation assay in bacteria, performed similarly to the OECD No.471 guideline, strains TA1535, TA1537, TA1538, TA98 and TA100 of S. typhimurium were exposed to Sodium Trifluoroacetate diluted in DMSO at concentrations of 0.1, 0.5, 1.0, 5.0 and 10 mg/plate in the presence and absence of mammalian metabolic activation (fraction of S9 from the liver of Sprague-Dawley male rats treated by intraperitoneal injection with Aroclor 1254 dissolved in maize oil). The method of direct incorporation was used in this study.

Sodium Trifluoroacetate was tested up to limit concentration recommended in the guideline (5 mg/plate).

The positive controls induced the appropriate responses in the corresponding strains.

Under the test conditions, the test item Sodium trifluoroacetate did not show mutagenic activity in the bacterial reverse mutation test with Salmonella typhimurium as there was no evidence of induced mutant colonies over background.

In this study Sodium Trifluoroacetate was tested instead of trifluoroacetic anhydride (TFAH) according to a reliable analogue approach in order to be free of the cytotoxic effect on bacteria due to the extreme acid pH of resulting Trifluoroacetic acid (TFA) (0.45) formed when TFAH is mixed to the aqueous culture medium.

Therefore, Trifluoroacetic anhydride is not considered as mutagenic in the bacterial reverse mutation test.

This study is considered as acceptable as it satisfied the main criteria of the OECD guideline No. 471.