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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
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 peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity Tests: V. Results from the Testing of 311 Chemicals
Author:
Errol Zeiger, Beth Anderson, Steve Haworth, Timothy Lawlor, and Kristien Mortelmans
Year:
1992
Bibliographic source:
Environmental and Molecular Mutagenesis Volume 19, Supplement 21: 2-141 (1992)

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Procedure as described by Haworth et al 1983 with slight modification
Principles of method if other than guideline:
Preincubation assay was performed to evaluate the mutagenic nature of the test compound 4-(2-Naphthylamino)phenol
GLP compliance:
not specified
Type of assay:
bacterial gene mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Details on test material:
- Name of test material: 4-(2-Naphthylamino)phenol
- Molecular formula: C16H13NO
- Molecular weight: 235.285 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: appr. 88%
- Impurities (identity and concentrations): No data
Specific details on test material used for the study:
- Name of test material: 4-(2-Naphthylamino)phenol
- IUPAC name: 4-(2-naphthylamino)phenol
- Molecular formula: C16H13NO
- Molecular weight: 235.285 g/mol
- Substance type: Organic
- Physical state: No data
- Purity: appr. 88%
- Impurities (identity and concentrations): 12%

Method

Target gene:
Histidine
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 100, TA1535, TA 1537, TA 97 and TA98
Details on mammalian cell type (if applicable):
Not applicable
Additional strain / cell type characteristics:
not specified
Metabolic activation:
with and without
Metabolic activation system:
10% and 30% hamster liver S9 and 10% and 30% rat liver S9
Test concentrations with justification for top dose:
0, 0.3, 1, 3, 10, 33, 66, 100, 166, 333 or 666 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: No data available
Controls
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylenediamine, 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: Preincubation

DURATION
- Preincubation period: 20 mins
- Exposure duration: 2 days
- Expression time (cells in growth medium): 2 days
- Selection time (if incubation with a selection agent): No data available
- Fixation time (start of exposure up to fixation or harvest of cells): No data available

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

NUMBER OF REPLICATIONS: No data available

NUMBER OF CELLS EVALUATED: No data available

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

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

OTHER: No data available
Rationale for test conditions:
No data
Evaluation criteria:
The plates were observed for a dose dependent increase in the number of Histidine- independent (his+) colonies.

Evaluations were made at both the individual trial and chemical levels.

Individual trials were judged mutagenic (+), weakly mutagenic (+ W), questionable (?), or nonmutagenic (-), depending on the magnitude of the increase in his+ revertants, and the shape of the dose response. A trial was considered questionable (?) if the dose-response was judged insufficiently high to support a call of “+ W”, if only a single dose was elevated over the control, or if a weak increase was not dose-related. The distinctions between a questionable response and a nonmutagenic or weakly mutagenic response, and between a weak mutagenic response and mutagenic response are highly subjective. It was not necessary for a response to reach two-fold over background for a trial to be judged mutagenic.

A chemical was judged mutagenic (+) or weakly mutagenic (+W) if it produced a reproducible, dose-related response over the solvent control, under a single metabolic activation condition, in replicate trials. A chemical was judged questionable (?) if the results of individual trials were not reproducible, if increases in his+ revertants did not meet the criteria for a “+W” response, or if only single doses produced increases in his+ revertants in repeat trials. Chemicals were judged nonmutagenic (-) if they did not meet the criteria for a mutagenic or questionable response.
Statistics:
Mean, SEM

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA 100, TA1535, TA 1537, TA 97 and TA98
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 available
Remarks on result:
other: No mutagenic potential

Any other information on results incl. tables

Table: Results for the test chemical 4-(2-Naphthylamino)phenol

Dose (µg/plate)

TA100

NA

10% HLI

30% HLI

10% RLI

30% RLI

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

0

91

5.4

89

6.2

139

5.6

102

9.0

154

4.4

0.3

125

5.8

 

 

 

 

 

 

 

 

1

104

10.0

 

 

 

 

 

 

 

 

3

91

2.9

82

7.3

 

 

98

6.7

 

 

10

85

0.7

105

21.6

134

15.7

108

17.0

181

8.9

33

46s

15.5

134

3.5

159

13.9

97

6.1

156

5.1

66

 

 

 

 

 

 

 

 

 

 

100

 

 

113

7.4

124

14.7

90

5.1

149

7.9

166

 

 

83

3.0

 

 

94

6.8

 

 

333

 

 

 

 

96

3.5

 

 

73

2.5

666

 

 

 

 

21s

9.4

 

 

0s

0.0

Positive control

352

7.2

681

36.9

612

23.9

414

7.2

550

31.4

 

 

Dose (µg/plate)

TA1535

NA

10% HLI

30% HLI

10% RLI

30% RLI

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

0

22

3.8

14

4.5

12

0.3

12

1.8

12

0.9

0.3

23

2.3

 

 

 

 

 

 

 

 

1.0

25

2.7

 

 

 

 

 

 

 

 

10

23

1.8

11

2.3

13

2.1

15

3.0

11

0.9

33

16

1.8

15

1.5

11

2.0

12

1.5

10

1.2

66

 

 

 

 

 

 

 

 

 

 

100

 

 

13

2.2

13

1.9

11

2.1

13

2.2

166

 

 

12

2.2

 

 

8

0.9

 

 

333

 

 

 

 

13

2.7

 

 

7

2.6

666

 

 

 

 

 

 

 

 

 

 

Positive control

301

18.6

269

22.6

400

31.3

165

9.2

122

17.9

 

Dose (µg/plate)

TA1537

NA

10% HLI

30% HLI

Mean

SEM

Mean

SEM

Mean

SEM

0

10

2.2

10

1.2

12

0.9

0.3

9

0.0

 

 

 

 

1

8

3.9

 

 

 

 

3

7

1.2

10

3.2

9

0.3

10

5

1.2

8

0.9

9

2.2

33

6

2.1

10

0.9

9

2.2

66

 

 

 

 

 

 

100

 

 

11

1.9

9

2.0

166

 

 

 

 

 

 

333

 

 

6

2.6

10

1.3

666

 

 

 

 

 

 

Positive control

397

27.1

46

4.0

61

0.6

 

Dose (µg/plate)

TA97

NA

10% HLI

30% HLI

10% RLI

30% RLI

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

0

189

9.6

190

20.5

167

12.3

193

10.8

174

11.7

0.3

198

2.3

 

 

 

 

 

 

 

 

1

195

3.5

 

 

 

 

 

 

 

 

3

202

7.1

205

8.0

173

13.6

193

0.7

174

4.9

10

175

12.0

194

5.0

156

14.3

204

9.0

186

12.3

33

184

12.1

170

3.5

201

7.0

208

4.5

182

13.3

66

 

 

 

 

 

 

 

 

 

 

100

 

 

121

13.0

189

9.3

190

16.5

159

23.1

166

 

 

67s

7.4

 

 

79

6.6

 

 

333

 

 

 

 

12

2.6

 

 

14

2.3

666

 

 

 

 

 

 

 

 

 

 

Positive control

433

9.6

332

11.3

343

18.3

316

6.7

351

12.2

 

Dose (µg/plate)

TA98

NA

10% HLI

30% HLI

10% RLI

30% RLI

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

0

14

2.6

27

2.2

23

3.0

34

3.1

24

1.3

0.3

17

3.5

 

 

 

 

 

 

 

 

1

18

3.8

 

 

 

 

 

 

 

 

3

16

2.5

37

3.4

 

 

28

1.5

 

 

10

14

0.6

39

0.3

31

5.5

21

2.7

24

1.2

33

13

1.5

31

2.5

25

3.0

15

2.3

22

1.5

66

 

 

 

 

 

 

 

 

 

 

100

 

 

22

1.7

21

1.7

19

2.5

17

1.0

166

 

 

21

3.5

 

 

18

1.5

 

 

333

 

 

 

 

12

1.8

 

 

12

1.3

666

 

 

 

 

9

0.9

 

 

6

1.2

Positive control

401

33.4

392

31.0

376

29.1

193

5.8

164

9.8

Applicant's summary and conclusion

Conclusions:
4-(2-Naphthylamino)phenol did not induce gene mutation in the Salmonella typhimurium tester strains TA 100, TA1535, TA 1537, TA 97 and TA98 in the presence and absence of S9 metabolic activation system and hence is negative for gene mutation in vitro.
Executive summary:

Preincubation assay was performed to evaluate the mutagenic nature of the 4-(2-Naphthylamino)phenol. The study was performed as described by Haworth et al 1983 with slight modification. The study involved the use of test chemical dose levels of 0, 0.3, 1, 3, 10, 33, 66, 100, 166, 333 or 666 µg/plate and Salmonella typhimurium strains TA 100, TA1535, TA 1537, TA 97 and TA98 in the presence and absence of S9 metabolic activation system. 4-(2-Naphthylamino)phenol did not induce gene mutation in the Salmonella typhimurium tester strains in the presence and absence of S9 metabolic activation system and hence is negative for gene mutation in vitro.