Bis(4-hydroxy-N-methylanilinium) sulphate

Genetic toxicity: in vitro

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

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

Test material information

Constituent 1

Reference substance name:

Bis(4-hydroxy-N-methylanilinium) sulphate

EC Number:

200-237-1

EC Name:

Bis(4-hydroxy-N-methylanilinium) sulphate

Cas Number:

55-55-0

Molecular formula:

C14H20N2O6S

IUPAC Name:

bis(4-hydroxy-N-methylanilinium) sulfate

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 / 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

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 / solvent:

- Vehicle(s)/solvent(s) used: DMSO- Justification for choice of solvent/vehicle: The chemical was soluble and stable 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:

9-aminoacridine

sodium azide

other: 2-Aminoanthracene (2-AA) (All strains; +S9), 4-nitro-o-phenylenediamine (TA98; -S9)

Details on test system and experimental 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 results

Test results 1

Species / strain:

S. typhimurium, other: TA98 and TA1535

Metabolic activation:

with and without

Genotoxicity:

negative

Remarks:

with 10% HLI or 10% RLI

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 2

Species / strain:

S. typhimurium, other: TA100 and TA97

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

Test results 3

Species / strain:

S. typhimurium, other: TA100

Metabolic activation:

with

Genotoxicity:

negative

Remarks:

from 10-1000 µg/plate, with 10% HLLI

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 4

Species / strain:

S. typhimurium TA 100

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

with 10% RLI

Cytotoxicity / choice of top concentrations:

not specified

Vehicle controls validity:

valid

Untreated negative controls validity:

not specified

Positive controls validity:

valid

Test results 5

Species / strain:

S. typhimurium TA 1537

Metabolic activation:

with

Genotoxicity:

positive

Remarks:

from 10-1667 µg/plate; with 10% HLI or 10% RLI

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:

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.