Registration Dossier

Toxicological information

Genetic toxicity: in vitro

Currently viewing:

Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
data from handbook or collection of data
Justification for type of information:
Data is from NTP.

Data source

Reference
Reference Type:
review article or handbook
Title:
Gene mutation toxicity study of the test chemical
Author:
National Institute of Environmental Health Sciences
Year:
2018
Bibliographic source:
National Toxicology Program (NTP), 2019

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
To evaluate the mutagenic potential of test chemical in Salmonella Typhimurium TA98,TA 100,TA 1535 and TA 97 by AMES test.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Trichloroacetic acid
EC Number:
200-927-2
EC Name:
Trichloroacetic acid
Cas Number:
76-03-9
Molecular formula:
C2HCl3O2
IUPAC Name:
2,2,2-Trichloroacetic acid
Test material form:
solid
Details on test material:
IUPAC name: 2,2,2-trichloroacetate
Mol. formula: C2HCl3O2
Molecular Weight: 163.3869 gm/mol
Smiles: C(C(=O)O)(Cl)(Cl)Cl
InChI: 1S/C2HCl3O2/c3-2(4,5)1(6)7/h(H,6,7)

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA98,TA 100,TA 1535 and TA 97
Metabolic activation:
with and without
Metabolic activation system:
RLI = induced male Sprague Dawley rat liver S9
HLI = induced male Syrian hamster liver S9
Test concentrations with justification for top dose:
0, 33, 100, 333, 1000, 3333, 10000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Distilled water
- Justification for choice of solvent/vehicle: The test substance is soluble in Distilled water
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Distilled water
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-Aminoanthracene: 1.0 ug/Plate : TA 100, TA 97, TA 98 (+S9); 2.0 ug/Plate: TA 100, TA 97, TA 98, TA 1535 (+S9); 5.0 ug/Plate: TA 100, TA 97, TA 98, TA 1535 (+S9); 10.0 ug/Plate : TA 1535 (+S9). 4-Nitro-O-Phenylenediamine: TA 98 (-S9)
Details on test system and experimental conditions:
NUMBER OF REPLICATIONS:
- Number of independent experiments : 3
Evaluation criteria:
If the substance under test is mutagenic, the chemical-treated plates will have a much greater number of colonies than the negative control plates. A positive response is a reproducible, dose-related increase in mutant colonies in any single strain, with or without the addition of S9 metabolic enzymes. While there is no minimum percentage of increase required for a result to be considered positive, a twofold increase in mutant colonies in a treated plate is usually considered to be a positive (mutagenic) response. An equivocal response is any increase that is not reproducible, not dose-related, or not high enough in magnitude to be considered positive. A negative response occurs when no increases in mutant colonies are seen in the cultures treated with the test chemical, compared with the control.
Statistics:
Mean or Mean ± Standard Error Mean.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA 100, TA 98, TA 97 and TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity
Vehicle controls validity:
valid
Remarks:
Water
Untreated negative controls validity:
not specified
True negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
No data
Remarks on result:
other: No mutagenic potential observed

Any other information on results incl. tables

Table 1: TA100

Dose (ug/Plate)

Metabolic activation

Without S9

Without S9

With 10% Rat S9

With 30% Rat S9

With 10% Hamster S9

With 30% Hamster S9

Vehicle Control

124±7.1

124±3.2

133±9.7

144±6.9

119±7.7

130±9.6

33.0

120±5.8

123±7.7

 

 

 

 

100.0

118±11.1

133±7.7

131±3.5

135±5.8

124±7.4

129±2.6

333.0

106±4.3

127±9.3

124±5.0

134±3.8

123±8.4

139±2.1

1000.0

114±10.3

116±1.3

126±9.9

133±4.3

127±2.1

127±9.5

3333.0

95±4.0

110±5.9

132±8.4

140±10.2

118±5.5

135±4.2

10000.0

 

 

119±6.3

78±6.1

117±3.8

110±9.7

Positive control

951±25.6

974±25.5

593±13.8

629±50.7

641±19.1

1022±22.5

 

Table 2: TA1535

Dose (ug/Plate)

Metabolic activation

Without S9

Without S9

With 10% Rat S9

With 30% Rat S9

With 10% Hamster S9

With 30% Hamster S9

Vehicle Control

8±0.9

19±2.5

23±2.5

13±1.2

18±3.0

10±1.2

33.0

 

19±0.3

 

 

 

 

100.0

8±0.7

17±2.0

19±0.3

10±0.9

21±2.0

9±0.3

333.0

7±1.5

17±2.2

15±0.7

14±2.6

22±3.5

9±0.6

1000.0

7±0.0

16±1.7

23±4.1

12±2.3

17±4.2

6±0.7

3333.0

5±0.3

16±1.5

23±1.2

11±0.6

18±2.0

9±1.8

10000.0

Toxic

 

19±0.9

9±0.3

14±0.3

9±2.0

Positive control

980±23.7

1017±44.8

209±10.3

134±9.8

232±21.0

170±3.1

 

Table 3: TA97

Dose (ug/Plate)

Metabolic activation

Without S9

Without S9

With 10% Rat S9

With 30% Rat S9

With 10% Hamster S9

With 30% Hamster S9

Vehicle Control

144±9.4

156±3.9

181±8.1

173±11.9

185±4.4

170±4.8

33.0

 

153±7.6

 

 

 

 

100.0

147±9.0

169±11.1

180±7.4

149±10.0

152±9.2

176±11.3

333.0

160±5.9

145±11.0

192±6.5

170±9.5

151±11.5

150±3.9

1000.0

172±3.0

169±2.3

180±8.9

172±10.5

182±8.2

165±2.0

3333.0

163±10.7

155±10.7

164±2.8

177±5.5

178±5.5

169±5.7

10000.0

Toxic

 

167±10.0

88±5.8

161±8.6

98±10.0

Positive control

491±14.6

499±6.6

462±20.5

544±17.2

590±16.6

605±4.9

 

Table 4: TA98

Dose (ug/Plate)

Metabolic activation

Without S9

Without S9

With 10% Rat S9

With 30% Rat S9

With 10% Hamster S9

With 30% Hamster S9

Vehicle Control

19±3.3

22±2.3

24±3.2

24±2.0

20±1.9

25±3.0

33.0

21±1.7

19±0.3

 

 

 

 

100.0

21±1.5

15±0.7

19±0.3

20±0.7

19±1.2

23±2.4

333.0

20±2.3

16±1.5

18±3.3

20±2.3

22±1.7

20±3.5

1000.0

23±2.6

21±4.2

26±2.4

23±1.3

22±1.0

21±1.5

3333.0

10±0.9

13±3.1

27±2.1

16±1.2

24±2.0

24±1.5

10000.0

 

 

21±1.7

9±0.9

20±2.7

10±1.5

Positive control

319±7.7

347±16.8

250±19.5

511±34.1

448±17.4

841±75.9

 

 

Applicant's summary and conclusion

Conclusions:
Test chemical was evaluated for its mutagenic potential in Salmonella typhimurium TA100, TA1535, TA98 and TA97 by AMES test. The test result was considered to be negative in all strain in the presence and absence of metabolic activation S9.

Executive summary:

Genetic toxicity in vitro study was assessed for test chemical. For this purpose AMES test was performed. The test material was exposed to Salmonella typhimurium TA100, TA1535, TA98 and TA97 in the presence and absence of metabolic activation S9. The S9 mix were prepared from 10% and 30% induced male Sprague Dawley rat liver S9 and induced male Syrian hamster liver S9 repectively. The concentration of test material used in the presence and absence of metabolic activation were 0, 33, 100, 333, 1000, 3333, 10000 µg/plate. No mutagenic effects were observed in any strains, in the presence and absence of metabolic activation. Therefore, test chemical was considered to be not mutagenic in Salmonella typhimurium TA100, TA1535, TA98 and TA97 by AMES test. Hence, the substance cannot be classified as gene mutant in vitro.