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Genetic toxicity: in vitro

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Administrative data

Endpoint:
genetic toxicity in vitro
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 J check

Data source

Reference
Reference Type:
review article or handbook
Title:
Gene mutation toxicity study of the test chemical
Author:
NITE
Year:
2017
Bibliographic source:
J-check

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: Isoamyl phenylacetate
- Molecular formula: C13H18O2
- Molecular weight: 183.05 g/mol
- Substance type: Organic
- Purity: 99.6%

Method

Target gene:
Histidine
Species / strainopen allclose all
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Species / strain / cell type:
E. coli WP2 uvr A
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Cytokinesis block (if used):
No data
Metabolic activation:
with and without
Metabolic activation system:
S9 mix
Test concentrations with justification for top dose:
All strains: 0, 1.22, 4.88, 19.5, 78.1, 313, 1250 or 5000 µg/plate

TA100:
Without S9: 0, 2.44, 4.88, 9.77, 19.5, 39.1, 78.1 µg/plate
With S9: 0, 9.77, 19.5, 39.1, 78.1, 156, 313 µg/plate

TA1535: With and without S9: 0, 2.44, 4.88, 9.77, 19.5, 39.1, 78.1 µg/plate

TA98: With and without S9: 0, 9.77, 19.5, 39.1, 78.1, 156, 313 µg/plate

TA1535: With and without S9: 0, 2.44, 4.88, 9.77, 19.5, 39.1, 78.1 µg/plate

E.coli WP2 uvrA: With and without S9: 0, 9.77, 19.5, 39.1, 78.1, 156, 313 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The chemical was soluble in DMSO
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
other: 2- (2- Furyl)-3-(5-nitro-2-furyl)acrylamide (TA100, WP2uvrA, TA98, -S9), 2-Methoxy-6-chloro-9-[3-(2-chloroethyl)-aminopropylamino]acridine.2HCl (TA1537, -S9), 2-Aminoanthracene (TA1535, WP2uvrA, +S9), Benzo[a]pyrene (TA100, TA1537, TA98, +S9)
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar

DURATION
- Preincubation period: No data
- Exposure duration: No data
- Expression time (cells in growth medium): No data
- Selection time (if incubation with a selection agent): No data
- Fixation time (start of exposure up to fixation or harvest of cells): No data

SELECTION AGENT (mutation assays): No data
SPINDLE INHIBITOR (cytogenetic assays): No data
STAIN (for cytogenetic assays): No data

NUMBER OF REPLICATIONS: Triplicate plates were used/dose level and the study was performed in triplicates

NUMBER OF CELLS EVALUATED: No data

DETERMINATION OF CYTOTOXICITY
- Method: mitotic index; cloning efficiency; relative total growth; other: No data

OTHER EXAMINATIONS:
- Determination of polyploidy: No data
- Determination of endoreplication: No data
- Other: No data

OTHER: No data
Rationale for test conditions:
No data
Evaluation criteria:
The plates were observed for a dose dependent increase in the number of revertants/plate
Statistics:
No data

Results and discussion

Test resultsopen allclose all
Species / strain:
other: TA100, TA1535, TA98, TA1535
Metabolic activation:
with and 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:
E. coli WP2 uvr A
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
not specified
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
No data
Remarks on result:
other: No mutagenic potential

Any other information on results incl. tables

Table 1:

Metabolic activation system

Dose (µg/plate)

TA100

TA1535

WP2uvrA

TA98

TA1537

Without S9

DMSO

98

11

18

27

10

113

11

17

32

7

 

4.88

119

1

15

26

7

19.5

112

5

15

25

6

78.1

109

7

15

23

8

313

0

0

0

0

0

1250

0

0

0

0

0

5000

0

0

0

0

0

With S9

DMSO

166

16

21

53

12

1.22

195

14

20

53

7

4.88

914

17

20

49

8

19.5

229

10

21

49

5

78.1

302

14

27

61

9

313

0

0

0

0

0

1250

0

0

0

0

0

5000

0

0

0

0

0

Positive control

 

AF-2

SAZ

AF-2

AF-2

ICR-191

 

535

331

81

436

1550

Positive control

 

BAP

2AA

2AA

BAP

BAP

 

835

299

1153

292

118

 

Table 2:

Metabolic activation system

Dose (µg/plate)

TA100

TA1535

WP2uvrA

TA98

TA1537

Without S9

DMSO

125±17.1

14±3.5

28±1.0

16±3.6

8±2.5

2.44

119±15.5

7±4.0

NT

NT

7±1.0

4.88

120±4.6

11±2.5

NT

NT

8±2.5

9.77

125±2.3

10±5.1

26±6.8

18±2.9

5±1.5

19.5

117±7.5

8±3.8

26±4.0

20±11.9

6±2.3

39.1

121±2.1

12±5.1

25±6.0

17±4.0

5±0.6

78.1

112±10.4

7±0.6

20±8.1

15±2.5

6±2.0

156

NT

NT

0±0.0

0±0.0

NT

313

NT

NT

0±0.0

0±0.0

NT

 

 

Table 3:

Metabolic activation system

Dose (µg/plate)

TA100

TA1535

WP2uvrA

TA98

TA1537

Without S9

DMSO

143±9.0

8±2.5

25±7.1

51±11.3

11±2.9

2.44

NT

13±2.1

NT

NT

14±3.0

4.88

NT

8±3.1

NT

NT

9±3.5

9.77

151±5.3

15±2.6

28±13.1

57±8.2

9±3.2

19.5

166±7.5

9±1.0

28±3.0

44±7.0

10±1.5

39.1

188±12.9

18±4.2

30±4.5

47±5.7

17±4.0

78.1

237±29.5

12±3.2

25±5.9

53±10.7

11±3.2

156

127±10.5

NT

29±6.2

31±2.9

NT

313

0 0.0

NT

0 0.0

0 0.0

NT

 

Table 4:

Metabolic activation system

Dose (µg/plate)

TA100

TA1535

WP2uvrA

TA98

TA1537

Without S9

DMSO

107±18.0

14±3.8

12±1.7

19±9.5

9±0.6

2.44

121±5.1

15±1.5

NT

NT

6±1.5

4.88

118±7.0

9±3.6

NT

NT

4±0.0

9.77

121±9.9

10±2.3

14±5.2

19±1.7

5±2.5

19.5

123±12.3

7±1.2

15±1.7

19±3.2

9±2.3

39.1

112±13.5

8±3.6

21±5.5

19±3.5

5±1.0

78.1

94±3.5

7±1.0

16±4.5

20±5.2

7±3.1

156

NT

NT

12±4.6

0±0.0

NT

313

NT

NT

0 0.0

0±0.0

NT

 

Table 5:

Metabolic activation system

Dose (µg/plate)

TA100

TA1535

WP2uvrA

TA98

TA1537

Without S9

DMSO

124±15.1

9±3.2

19±6.0

46±7.5

11±2.6

2.44

NT

9±2.0

NT

NT

9±3.5

4.88

NT

8±1.5

NT

NT

7±1.2

9.77

142±11.4

9±1.2

15±1.0

39±3.1

8±2.9

19.5

151±13.3

8±3.8

16±3.0

42±7.6

10±2.5

39.1

186±12.7

16±5.0

16±4.7

47±13.1

10±2.1

78.1

212±13.3

11±4.0

16±2.6

44±6.1

11±2.6

156

115±22.1

NT

17±3.2

27±4.6

NT

313

0±0.0

NT

0±0.0

0±0.0

NT

Applicant's summary and conclusion

Conclusions:
The test chemical did not induce gene mutation in Salmonella typhimurium strains TA100, TA1535, TA98, TA1535 and E. coli WP2 uvrA in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.
Executive summary:

Gene mutation toxicity study was performed to determine the mutagenic nature of the test chemical. The study was performed as per the OECD guideline 471 using Salmonella typhimurium strains TA100, TA1535, TA98, TA1535 and E. coli WP2 uvrA with and without of S9 metabolic activation system. The study was performed at various dose levels and in triplicate plates/dose. Concurrent solvent and positive control chemicals were incorporated in the study. The plates were observed for dose dependent increase in the number of revertnats/plate. The test chemical did not induce gene mutation in Salmonella typhimurium strains TA100, TA1535, TA98, TA1535 and E. coli WP2 uvrA in the presence and absence of S9 metabolic activation system and hence is not likely to classify as a gene mutant in vitro.