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Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Salmonella Mutagenicity Tests of test chemical was performed in Salmonella strain TA98, TA100, TA1535, TA 1537 and TA97 in the presence and absence of S9 metabolic activation system produced mutation, therefore it is considered to be positive for gene mutation in vitro.

Link to relevant study records
Reference
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:
data from handbook or collection of data
Justification for type of information:
Data is from publication
Qualifier:
according to
Guideline:
other: As mentioned below
GLP compliance:
no
Type of assay:
bacterial reverse mutation assay
Target gene:
Histidine
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
10%HLI and 10%RLI

The S-9 (9000g supernatant) fractions of Aroclor 1254-induced, male Sprague- Dawley rat and male Syrian hamster livers were prepared. The S-9 mixes were prepared immediately prior to use and contained either 10% or 30% S-9; occasionally, other levels were used.

Test concentrations with justification for top dose:
Dose :
0, 33, 100, 333, 1000 and 1666 µg/plate
Vehicle / solvent:
Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was soluble in DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Dimethyl Sulfoxide
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
Control for- test without S9 activation (TA 1535 and TA 100)
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Dimethyl sulfoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
Absence of metabolic activation (TA 97 and TA 1537)
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Dimethyl sulfoxide
True negative controls:
no
Positive controls:
yes
Positive control substance:
other: 4-nitro-o-phenylenediamine
Remarks:
With metabollic activation (TA 98)
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
Dimethyl Sulfoxide
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
other: 2-aminoanthracene
Remarks:
Control for- test with HLI and RLI S9 activation
Rationale for test conditions:
Not specified
Evaluation criteria:
A chemical was judged mutagenic (+) or weakly mutagenic (+ W) if it produced a reproducible dose-related reponse over the solvent control in replicate trials.
The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or nonmutagenicity. The plate count means are presented in Appendix 2 so that the reader has the opportunity of performing his or her own evaluation of the data.
Statistics:
Not specified
Species / strain:
other: S. typhimurium TA 1535, 100, 97, 98
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
other: S. typhimurium TA 1537, 100, 98
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
other: S. typhimurium TA 1535, 1537, 100, 97
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
other: S. typhimurium TA 1535, 97
Metabolic activation:
with
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with
Genotoxicity:
positive
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: all strains/cell types tested

Strain: TA 100

Dose

NA

(-)

 

10% HLI

(+)

 

10% RLI

(+W)

 

µg/plate

Mean

SEM

Mean

SEM

Mean

SEM

0.000

152

13.9

173

2.4

143

15.5

33.000

137

18.8

271

21.4

194

3.8

100.000

133

2.9

301

4.1

213

2.9

333.000

116

6.4

409

14.1

254

3.4

1000.000

139

10.8

451

9.0

265

13.7

1666.000

23S

5.0

446

17.0

235

13.2

POS

500

4.1

1007

31.3

953

15.3

               

Strain:TA1537

Dose

10% HLI

(+)

 

10% RLI

(-)

 

µg/plate

Mean

SEM

Mean

SEM

0.000

5

1.7

12

0.3

33.000

9

2.8

9

0.9

100.000

11

1.5

12

1.7

333.000

19

3.3

13

1.5

1000.000

25

0.7

17

0.9

1666.000

43

4.6

14

0.7

POS

322

21.1

153

5.0

 

 

Strain:TA97

Dose

NA

(?)

 

10% HLI

(+W)

 

10% RLI

(-)

 

µg/plate

Mean

SEM

Mean

SEM

Mean

SEM

0.000

118

9.3

196

3.5

183

16.9

33.000

181

4.4

214

17.8

178

2.5

100.000

187

5.9

213

3.5

179

12.5

333.000

179

4.7

264

6.0

158

6.1

1000.000

157

13.1

272

17.7

187

16.2

1666.000

81S

12.3

290

21.0

82S

21.2

POS

652

9.1

777

23.3

638

5.9

 

 

 

Strain:TA98

Dose

NA

(-)

 

10% HLI

(+)

 

10% RLI

(+)

 

µg/plate

Mean

SEM

Mean

SEM

Mean

SEM

0.000

30

1.2

42

3.5

40

0.0

33.000

28

4.2

46

2.8

36

4.1

100.000

31

2.8

59

3.0

46

2.7

333.000

25

3.1

105

4.1

67

3.2

1000.000

24

5.3

187

8.2

97

15.0

1666.000

5S

1.3

292

4.5

92

8.1

POS

814

36.0

549

23.5

403

37.4

Strain : TA 1535

Dose No Activation
(Negative)
10% HLI
(Negative)
10% RLI
(Negative)
Protocol Preincubation Preincubation Preincubation
ug/Plate Mean ± SEM Mean ± SEM Mean ± SEM
0     
34 3.3 12 1.2 13 1.5
33     
35 1.9 11 1.3 14 0.9
100     
36 4.9 10 1.9 10 1.5
333     
30 0.3 15 0.3 12 2
1000     
36 2.6 13 0.3 9 0.9
1666     
2s 0.6 10 0.6 12 2.1
Positive Control 468 11.5 269 9.5 245 15.9
Abbreviations:
RLI = induced male Sprague Dawley rat liver S9
HLI = induced male Syrian hamster liver S9
s = Slight Toxicity; p = Precipitate; x = Slight Toxicity and Precipitate; T = Toxic; c = Contamination
Conclusions:
Salmonella Mutagenicity Tests of test chemical was performed in Salmonella strain TA98, TA100, TA1535, TA 1537 and TA97 in the presence and absence of S9 metabolic activation system produced mutation, therefore it is considered to be positive for gene mutation in vitro.
Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using Salmonella typhimurium strains TA98, TA97, TA100, TA1537 and TA1535 in the presence and absence of 10% Hamster Liver Infusion S9 and 10% Rat Liver Infusion S9 metabolic activation system. The chemical was dissolved in DMSO as solvent and used at dose levels 0, 33.0, 100.0, 333.0, 1000.0 and 1666.0 µg/plate by the preincubation method. Sodium azide, 9-aminocridine, 4-nitro-o-phenylenediamine and 2-aminoanthracene were used as positive control substances in the presence and absence of metabolic activation.

Genetic toxicity of test chemical to Salmonella typhimurium strain TA1535, TA100, TA97 and TA 98 was observed to be negative without S9 metabolic activation. Salmonella typhimurium strain TA1537, TA100 and TA 98 was observed to be negative with hamster, liver, S-9, aroclor 1254 (10%) induced metabolic activation. Salmonella typhimurium Strain TA1537, TA100, TA 1535 and TA 97 was observed to be negative with rat, liver, s-9, aroclor 1254 (10%) induced metabolic activation.

From the above observations it can be concluded that the test chemical is weakly mutagenic in nature.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed (positive)

Additional information

Data available for the target chemical was reviewed to determine the mutagenic nature of the test chemical. The studies are as mentioned below:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using Salmonella typhimurium strains TA98, TA97, TA100, TA1537 and TA1535 in the presence and absence of 10% Hamster Liver Infusion S9 and 10% Rat Liver Infusion S9 metabolic activation system. The chemical was dissolved in DMSO as solvent and used at dose levels 0, 33.0, 100.0, 333.0, 1000.0 and 1666.0 µg/plate by the preincubation method. Sodium azide, 9-aminocridine, 4-nitro-o-phenylenediamine and 2-aminoanthracene were used as positive control substances in the presence and absence of metabolic activation.

Genetic toxicity of test chemical to Salmonella typhimurium strain TA1535, TA100, TA97 and TA 98 was observed to be negative without S9 metabolic activation. Salmonella typhimurium strain TA1537, TA100 and TA 98 was observed to be negative with hamster, liver, S-9, aroclor 1254 (10%) induced metabolic activation. Salmonella typhimurium Strain TA1537, TA100, TA 1535 and TA 97 was observed to be negative with rat, liver, s-9, aroclor 1254 (10%) induced metabolic activation.

From the above observations it can be concluded that the test chemical is weakly mutagenic in nature.

In another study, Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed using Salmonella typhimurium strains TA98, TA100 and TA1537 in the presence and absence of Rat Liver Infusion S9 metabolic activation system. The chemical was dissolved in DMSO as solvent and used at dose levels 0.1 ml by the salmonella/microsome method.

Salmonella typhimurium strains TA98 and TA1537 did not show mutagenic nature in the absence of S9 metabolic activation. Whereas, 58 revertants /µmole/plate were observed in the agar plate and weakly positive results were observed in Salmonella typhimurium strain TA100 in the he presence of S9 metabolic activation system. Therefore the test chemical is considered to be mutagenic in nature.

Based on the data available for the test chemical, the test chemical induced mutation in the Salmonella typhimurium strain TA98, TA97, TA100, TA1535 or TA1537 both in the presence and absence of S9 metabolic activation system and hence is likely to be mutagenic under the conditions of this study.

Justification for classification or non-classification

Based on the observations made, the test chemical exibits gene mutation in vitro . Hence it is likely to classify as a gene mutant in vitro as per the criteria mentioned in CLP regulation.