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:
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 peer reviewed publication

Data source

Reference
Reference Type:
publication
Title:
Salmonella Mutagenicity Tests: III. Results from the Testing of 255 Chemicals
Author:
Errol Zeiger, Beth Anderson, Steve Haworth, Timothy Lawlor, Kristien Mortelmans, and William Speck
Year:
1987
Bibliographic source:
Environmental Mutagenesis Volume 9, Supplement 9: 1-110,1987

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Principles of method if other than guideline:
A bacterial reverse mutation assay was performed to evaluate the mutagenic nature of the test compound N-Methyl-p-aminophenol sulfate.
GLP compliance:
not specified
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: crystalline
Details on test material:
- Name of test material: p-Methylaminophenol sulfate- IUPAC name: Bis(4-hydroxy-N-methylanilinium) sulphate- Molecular formula: C14H20N2O6S- Molecular weight: 344.386 g/mole- Smiles:CNc1ccc(cc1)O.CNc1ccc(cc1)O.OS(=O)(=O)O- Inchl: 1S/2C7H9NO.H2O4S/c2*1-8-6-2-4-7(9)5-3-6;1-5(2,3)4/h2*2-5,8-9H,1H3;(H2,1,2,3,4)- Substance type: Organic- Physical state: Solid crystalline (off white - white)
Specific details on test material used for the study:
- Name of test material: N-Methyl-p-aminophenol sulfate- IUPAC name: 4-(Methylamino)phenol Sulfate - Molecular formula: C7H9NO.1/2H2O4S- Molecular weight: 344.386 g/mol- Substance type: Organic- Physical state: No data - Purity: 99+%- Impurities (identity and concentrations): No data

Method

Target gene:
Histidine
Species / strain
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Details on mammalian cell lines (if applicable):
Not applicable
Additional strain characteristics:
not specified
Cytokinesis block (if used):
No data available
Metabolic activation:
with and without
Metabolic activation system:
The S-9 fractions of Aroclor 1254-induced, male Sprague-Dawley rat and male Syrian hamster livers were prepared
Test concentrations with justification for top dose:
0, 0.33, 1.0, 3.3, 10, 33.0, 100, 167, 333, 667, 1000 or 1667 µg/plate
Vehicle:
- Vehicle(s)/solvent(s) used: DMSO- Justification for choice of solvent/vehicle: The chemical was soluble and stable in DMSO
Controls
Negative controls:
not specified
Solvent controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 2-Aminoanthracene (2-AA) (All strains; +S9), 4-nitro-o-phenylenediamine (TA98; -S9)
Details on test system and conditions:
METHOD OF APPLICATION: PreincubationDURATION- Preincubation period: 20 mins- Exposure duration: 48 hrs- Expression time (cells in growth medium): 48 hrs- 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 availableSELECTION AGENT (mutation assays): No data availableSPINDLE INHIBITOR (cytogenetic assays): No data availableSTAIN (for cytogenetic assays): No data availableNUMBER OF REPLICATIONS: TriplicateNUMBER OF CELLS EVALUATED: No data availableDETERMINATION OF CYTOTOXICITY- Method: mitotic index; cloning efficiency; relative total growth; other: No data availableOTHER EXAMINATIONS:- Determination of polyploidy: No data available- Determination of endoreplication: No data available- Other: No data availableOTHER: No data available
Rationale for test conditions:
No data available
Evaluation criteria:
An individual trial was judged mutagenic (+) if a dose-related increase over the corresponding solvent control was seen, and it was judged weakly mutagenic C+W) if a low-level dose response was seen. A trial was considered questionable (?) if a dose-related increase was judged insufficiently high to justify a call of "+W," if only a single dose was elevated over the control, or if a non-dose-related increase was seen. The chemical was judged to be mutagenic (+), or weakly mutagenic (+W), if it produced a reproducible, dose-related increase in his+ revertants over the corresponding solvent controls in replicate trials. A chemical was considered to be questionable (?) if a reproducible increase of his+ revertants did not meet the criteria for either a " + " or " + W," or if only single doses produced an increase in his+ revertants in repeat trials.
Statistics:
No data available

Results and discussion

Test resultsopen allclose all
Species / strain:
other: TA98 and TA1535
Metabolic activation:
with and without
Genotoxicity:
negative
Remarks:
with 10% HLI or 10% RLI
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Species / strain:
other: TA100 and TA97
Metabolic activation:
without
Genotoxicity:
negative
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Species / strain:
other: TA100
Metabolic activation:
with
Genotoxicity:
negative
Remarks:
from 10-1000 µg/plate, with 10% HLLI
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with
Genotoxicity:
positive
Remarks:
with 10% RLI
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with
Genotoxicity:
positive
Remarks:
from 10-1667 µg/plate; with 10% HLI or 10% RLI
Cytotoxicity:
not specified
Vehicle controls valid:
yes
Negative controls valid:
not specified
Positive controls valid:
yes
Additional information on results:
TEST-SPECIFIC CONFOUNDING FACTORS- Effects of pH: No data available- Effects of osmolality: No data available- Evaporation from medium: No data available- Water solubility: No data available- Precipitation: No data available- Other confounding effects: No data availableRANGE-FINDING/SCREENING STUDIES: The chemical was tested initially in a toxicity assay to determine the appropriate dose range. The toxicity assay was performed by using TA100 or the system developed by Waleh et al. Toxic concentrations were those at which a decrease in the number of his+ colonies was seen or at which there was a clearing in the density of the background lawn.COMPARISON WITH HISTORICAL CONTROL DATA: No data available ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Any other information on results incl. tables

Table: Mutagenicity of N-Methyl-p-aminophenol sulfate

Dose (µg/plate)

TA100

-S9

-S9

10% HLI

10% RLI

10% HLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

Mean

SEM

0

115

5.2

111

3.5

179

9.8

148

18.2

137

6.7

144

9.3

0.3

 

 

119

7.8

 

 

 

 

 

 

 

 

1.0

117

7.9

114

0.6

 

 

 

 

 

 

 

 

3.3

124

1.9

119

1.3

 

 

 

 

 

 

 

 

10

146

3.8

128

9.3

187

6.8

143

6.4

181

6.5

120

6.7

33

108

2.0

130

18.7

163

2.3

132

2.8

147

4.0

148

10.3

100

0

0.0

 

 

160

13.7

125

3.6

168

3.5

161

7.7

167

 

 

 

 

 

 

 

 

 

 

 

 

333

 

 

 

 

253

4.5

158

11.6

209

24.8

198

10.7

667

 

 

 

 

 

 

 

 

 

 

 

 

1000

 

 

 

 

217

23.6

163

8.4

2128

6.4

208

11.1

1667

 

 

 

 

 

 

 

 

 

 

 

 

Positive control

559

39.4

447

20.9

1903

31.3

1908

157.3

965

51.5

2338

40.7

Dose (µg/plate)

TA1535

-S9

10% HLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

0

6

1.2

5

1.2

6

0.5

0.3

4

0.6

 

 

 

 

1.0

5

1.3

 

 

 

 

3.3

3

1.0

 

 

 

 

10

3

0.3

6

2.5

5

0.9

33

5

0.6

6

2.2

8

2.5

100

 

 

7

1.3

6

1.5

167

 

 

 

 

 

 

333

 

 

5

1.2

6

1.2

667

 

 

 

 

 

 

1000

 

 

T

 

6

2.3

1667

 

 

 

 

 

 

Positive control

154

16.5

38

3.5

39

8.5

 

Dose (µg/plate)

TA98

-S9

10% HLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

0

13

2.3

14

1.7

12

1.0

0.3

10

1.5

 

 

 

 

1.0

8

2.1

 

 

 

 

3.3

17

11.3

 

 

 

 

10

11

3.3

19

3.0

13

2.1

33

T

 

16

2.4

13

3.0

100

 

 

14

3.0

16

2.2

167

 

 

 

 

 

 

333

 

 

16

2.6

14

1.3

667

 

 

 

 

 

 

1000

 

 

12

5.5

15

2.0

1667

 

 

 

 

 

 

Positive control

1668

21.1

938

65.1

1670

94.1

 

Dose (µg/plate)

TA1537

-S9

-S9

10% HLI

10% HLI

10% HLI

10% RLI

10% RLI

10% RLI

Mean

SEM

Mean

SEM

Mean

SEM

Mean

Mean

Mean

Mean

Mean

SEM

Mean

SEM

Mean

SEM

0

4

0.6

5

0.7

5

0.9

3

0.9

5

1.2

6

1.7

5

1.2

5

0.3

0.3

 

 

6

2.2

 

 

 

 

 

 

 

 

 

 

 

 

1.0

3

0.3

7

2.6

 

 

 

 

 

 

 

 

 

 

 

 

3.3

3

0.6

6

1.5

 

 

 

 

 

 

 

 

 

 

 

 

10

3

0.3

6

1.7

6

0.3

3

1.0

 

 

8

0.3

5

1.5

 

 

33

T

 

7

0.9

13

0.7

6

1.7

 

 

6

0.6

4

1.0

 

 

100

0

0.0

 

 

11

0.9

5

1.5

 

 

10

1.5

7

2.2

 

 

167

 

 

 

 

 

 

 

 

14

3.0

 

 

 

 

15

1.0

333

 

 

 

 

21

1.9

15

2.6

22

3.3

6

1.5

9

2.6

11

1.0

667

 

 

 

 

 

 

 

 

26

5.5

 

 

 

 

18

0.3

1000

 

 

 

 

20

1.9

20

3.5

T

 

22

0.9

23

1.3

19

3.1

1667

 

 

 

 

 

 

 

 

0

0.0

 

 

 

 

0

0.0

Positive control

308

26.2

644

56.9

340

26.2

109

13.5

1203

272.8

39

1.5

59

4.2

267

56.3

T, complete clearing of background lawn (colonies not counted)

Applicant's summary and conclusion

Conclusions:
N-Methyl-p-aminophenol sulfate did not induce a reproducible, dose-related increase in his+ revertants over the corresponding solvent in the S. typhimurium tester strains TA1535 and TA98 in the presence and absence of S9 metabolic activation system. It was also non mutagenic for the strains TA100 (10-1000 µg/plate) in the presence of metabolic activation system from hamster and TA1537 in the absence of S9 metabolic activation system. However, it induced gene mutation in the strains TA 100 (with S9 from rat) and TA1537 in the presence of S9 from 10-1667 µg/plate.
Executive summary:

A Salmonella/microsome test was performed in the presence and absence of exogenous metabolic activation by S9-frqactions from the livers of Aroclor-induced male Sprague-Dawley rats or Syrian hamsters to evaluate the mutagenic nature of N-Methyl-p-aminophenol sulfate in Salmonella typhimurium tester strains TA98, TA100, TA1535 and TA1537. The study was performed as per the preincubation assay with a preincubation time of 20 mins and then an incubation for 48 hrs. The test compound was administered at a dosage level of 0, 0.3, 1.0, 3.3, 10, 33.0, 100, 167, 333, 667, 1000 or 1667 µg/plate and concurrent solvent and positive control chemicals were included in the study. N-Methyl-p-aminophenol sulfate did not induce a reproducible, dose-related increase in his+ revertants over the corresponding solvent in the S. typhimurium tester strains TA1535 and TA98 in the presence and absence of S9 metabolic activation system. It was also non-mutagenic for the strains TA100 (10-1000 µg/plate) in the presence of metabolic activation system from hamster and TA1537 in the absence of S9 metabolic activation system. However, it induced gene mutation in the strains TA100 (combined with S9 factions from rats) and TA1537 in the presence of S9 from 10-1667 µg/plate. Therefore, the results indicate that a mutagenic response can be induced in vitro and thus the results are interpret as ambiguous.