N,N-diethylaniline

Genetic toxicity: in vitro

Administrative data

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 peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Gene mutation toxicity study of the test chemical

Author:

Zeiger et al

Year:

1988

Bibliographic source:

Environmental and Molecular Mutagenesis

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:

Preincubation method was performed to determine the mutagenic nature of the test chemical

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

N,N-diethylaniline

EC Number:

202-088-8

EC Name:

N,N-diethylaniline

Cas Number:

91-66-7

Molecular formula:

C10H15N

IUPAC Name:

N,N-diethylaniline

Details on test material:

- Name of test material: N,N-Diethylaniline
- Molecular formula: C10H15N
- Molecular weight: 149.236 g/mol
- Substance type: organic
- Physical state: solid
- Purity: 99%

Method

Target gene:

Histidine

Species / strain

Species / strain / cell type:

S. typhimurium, other: TA100, TA1535, TA97 and TA98

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:

10% and 30% HLI and RLI S-9 (9,000 g supernatant) fractions were prepared from Aroclor 1254-induced, male Sprague- Dawley rat and male Syrian hamster livers

Test concentrations with justification for top dose:

0, 1, 3, 10, 33, 100, 333 µg/plate

Vehicle / solvent:

Dimethyl Sulfoxide- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The test chemical was dissolved in DMSO

Controls

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

Remarks:

Dimethyl Sulfoxide

True negative controls:

not specified

Positive controls:

yes

Remarks:

in the absence of S9 activation

Positive control substance:

9-aminoacridine

sodium azide

other: 4-nitro-o-phenylenediamine (TA98, with S9) ; The positive control for metabolic activation with all strains was 2-aminoanthracene.

Details on test system and experimental conditions:

METHOD OF APPLICATION: preincubation

DURATION
- 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 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: Triplicate

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

Evaluation criteria:

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

Individual trials were judged mutagenic (+), weakly mutagenic (+ W), questionable
(?), or nonmutagenic (-), depending on the magnitude of the increase of 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 the increase seen was not dose related. The distinctions between a questionable mutagenic response and a nonmutagenic or weak 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 chemical to be judged mutagenic.

A chemical was judged mutagenic (+) or weakly mutagenic (+ W) if it produced a reproducible dose-related reponse over the solvent control 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. The chemicals were decoded by the chemical repository only after a determination had been made regarding their mutagenicity or nonmutagenicity.

Statistics:

Mean ± SD

Results and discussion

Test results

Species / strain:

S. typhimurium, other: TA100, TA1535, TA97 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:

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 available

RANGE-FINDING/SCREENING STUDIES: All chemicals were tested initially in a toxicity assay to determine the appropriate dose range for the mutagenicity assay. The toxicity assay was performed using TA100. Toxic concentrations were those that produced a decrease in the number of his+ colonies, or a clearing in the density of the background lawn, or both.

COMPARISON WITH HISTORICAL CONTROL DATA: Yes

ADDITIONAL INFORMATION ON CYTOTOXICITY: No data available

Remarks on result:

other: No mutagenic potential

Any other information on results incl. tables

Table: Mutagenicity of the test chemical

 

Dose (µg/plate)	TA100
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	127	7.0	98	3.3	129	3.6	93	1.2	147	6.2
1	119	6.6
3	116	11.7	96	2.6	134	7.9	103	6.7	119	8.3
10	96	7.9	96	10.2	124	4.3	103	6.1	132	10.5
33	123	10.0	97	6.5	143	4.4	104	5.0	123	7.1
100	127	14.3	97	5.0	131	8.5	97	7.0	123	8.1
333			52	226.2	136	10.6	80	4.5	112	6.4
Positive control	375	12.3	1828	130.2	873	46.0	725	24.1	449	7.0

 

Dose (µg/plate)	TA1535
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	24	3.4	11	3.3	12	2.4	10	4.0	12	6.0
1	23	4.4
3	26	3.8	14	5.0	10	2.9	10	1.5	12	3.5
10	18	1.2	9	1.8	9	3.2	7	1.7	22	1.5
33	22	4.3	10	2.2	13	0.6	11	2.7	14	2.0
100	12	0.7	7	1.9	7	1.5	10	1.5	12	1.5
333			4	0.3	15	1.9	7	2.7	15	2.8
Positive control	418	23.1	557	10.3	616	69.9	205	12.4	162	27.5

 

Dose (µg/plate)	TA97
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	121	8.1	117	5.5	234	15.2	193	8.2	236	9.4
1	134	0.3
3	108	7.8	133	4.4	204	12.1	169	10.3	207	21.5
10	129	637	143	5.8	206	15.2	156	10.5	211	4.6
33	132	11.2	138	6.4	207	10.7	139	7.1	223	13.0
100	108	2.0	144	6.1	197	10.2	123	9.0	186	23.8
333			113	6.0	186	9.2	129	18.0	222	14.5
Positive control	1156	22.0	1885	76.9	61.9	1452	80.7	480	60.6	7.0

 

Dose (µg/plate)	TA1535
	-S9		10% HLI		30% HLI		10% RLI		30% RLI
	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM	Mean	SEM
0	20	2.6	26	4.7	28	2.5	28	2.9	33	2.8
1	16	5.0
3	16	2.6	26	3.2	27	1.7	24	3.6	25	3.5
10	17	2.9	31	4.5	31	3.8	26	1.9	23	3.8
33	22	3.6	35	3.2	27	1.9	30	0.3	25	2.6
100	12	1.7	31	9.0	27	3.3	32	2.5	26	2.4
333			30	0.9	29	1.5	23	3.5	28	3.9
Positive control	845	69.2	1187	29.6	576	116.6	408	20.3	246	18.1

 

Applicant's summary and conclusion

Conclusions:

The test chemical is not mutagenic to the Salmonella typhimurium TA100, TA1535, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.

Executive summary:

The test chemical was studied for its ability to induce mutations in strains of Salmonella typhimurium.

 

The test compound was dissolved in DMSO and was tested at concentration of 0, 1, 3, 10, 33, 100 or 333 µg/plate in lab using Salmonella typhimurium TA100, TA1535, TA97 and TA98 in the presence and absence of 10 % and 30 % rat and hamster liver S9 metabolic activation system. Preincubation assay was performed with a preicubation for 20 mins. The plates were observed for histidine independence after 2 days incubation period. Concurrent solvent and positive controls were included in the study.

The test chemical is not mutagenic to the Salmonella typhimurium TA100, TA1535, TA97 and TA98 in the presence and absence of rat and hamster liver S9 metabolic activation system and hence it is not likely to classify as a gene mutant in vitro.