N-benzyl-N-ethylaniline

Endpoint summary

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Gene mutation toxicity study was performed by Errol Zeiger et al (Environmental and Molecular Mutagenesis, 1988) to determine the mutagenic nature of N-ethyl-N phenyl benzyl amine (92-59-1) using Salmonella typhimurium strains. In vitro genetic toxicity test for N-ethyl-N phenyl benzyl amine was performed on Salmonella typhimurium strains TA97, TA98, TA100, and TA1535 by Preincubation method. The test chemical (0.05 ml), Salmonella culture (0.10 ml), and S-9 mix or buffer (0.50 ml) was incubated at 37°C, without shaking, for 20 min. After that histidine-independent (his+) colonies were counted on plates by machine unless there was any precipitate was present which interfered with the count, or the colour of the test chemical on the plate reduced the contrast between the colonies and the background agar. The test material was exposed at the concentration of 0.000, 100.000, 333.000, 1000.000, 3333.000, 6666.000, 10000.000 µg/plate. There are some variations in protocol as the first test of a chemical was without activation and with 10% S-9 in the S-9 mix. If a positive result was obtained the test was repeated. If the tests were negative they were repeated without S-9 and with 30% S-9.Concurrent solvent and positive controls were run with each test. After exposure no mutagenic effect were observed. Therefore N-ethyl-N phenyl benzyl amine (92-59-1) was considered to be non mutagenic in Salmonella typhimurium strains TA97, TA98, TA100, and TA1535in the presence and absence of metabolic activation.

Link to relevant study records

Reference

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:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Justification for type of information:

Data is from publication .

Qualifier:

according to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

To evaluate the mutagenic potential of N-Ethyl-N-phenyl benzyl amine in Salmonella typhimurium strains TA97, TA98, TA100, and TA1535 by Ames tests.

GLP compliance:

not specified

Type of assay:

bacterial reverse mutation assay

Specific details on test material used for the study:

- Name of test material: N-benzyl-N-ethylaniline
- Molecular formula: C15H17N
- Molecular weight : 211.306 g/mol
- Smiles notation: N(c1ccccc1)(Cc1ccccc1)CC
- InChl: 1S/C15H17N/c1-2-16(15-11-7-4-8-12-15)13-14-9-5-3-6-10-14/h3-12H,2,13H2,1H3
- Substance type: Organic
- Physical state: Liquid

Target gene:

Histidine

Species / strain / cell type:

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

Details on mammalian cell type (if applicable):

Not applicable

Additional strain / cell type characteristics:

not specified

Cytokinesis block (if used):

Not specified .

Metabolic activation:

with and without

Metabolic activation system:

S-9 from Aroclor-induced male Sprague-Dawley rats and Syrian hamsters liver

Test concentrations with justification for top dose:

0.000,100.000,333.000,1000.000,3333.000,6666.000,10000.000 µg/plate

Vehicle / solvent:

DMSO(Dimethyl sulfoxide)

Untreated negative controls:

not specified

Negative solvent / vehicle controls:

yes

True negative controls:

not specified

Positive controls:

yes

Positive control substance:

other: Positive control in absence of metabolic activation: sodium azide (for strain TA1535 and TA100), 9-aminoacridine (TA97 and TA 1537), and 4-nitro-o-phenylenediamine (TA98). Positive control in presences of metabolic activation: 2-aminoanthracene

Details on test system and experimental conditions:

Details on test system and conditions
METHOD OF APPLICATION: Preincubation method

DURATION
- Preincubation period: 20 min
- Exposure duration: 2 days
- Expression time (cells in growth medium): 2 days

Rationale for test conditions:

Not specified

Evaluation criteria:

Number of histidine-independent (his+) colonies

Statistics:

Standard deviation was observed.

Key result

Species / strain:

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

Additional information on results:

RANGE-FINDING/SCREENING STUDIES: Chemical was tested by their half life dose intervals up to doses that elicited toxicity, or to a dose immediately below one which was toxic in the preliminary toxicity test and subsequent dose increments and may not have included doses in the toxic range were used.

N-benzyl-N-ethylaniline Result for AMES test

Dose (µg/plate)	TA100
	NA (-)		10% HLI (-)		10% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	103	3.5	105	9.4	128	6.4
100	94	2.9	98	8.6	107	2.4
333	95	1.9	95	3.2	127	5.9
1000	96	4.3	92	5.2	115	8.4
3333	90P	3.5	88P	0.9	98P	1.5
6666	89P	2.8
10000			100P	1.7	110P	1.9
Positive control	601	12.4	2276	47.7	724	34.5

 

Dose (µg/plate)	TA100
	NA (-)		30% HLI (-)		30% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	103	3.5	99	1.9	120	10
100	94	2.9	114	7.0	130	13
333	95	1.9	105	9.3	126	7.3
1000	96	4.3	97	3.8	105	7.2
3333	90P	3.5	88P	1.0	95P	8.7
6666	89P	2.8	-	-	-	-
10000			93P	3.8	88P	6.1
Positive control	601	12.4	2067	38.7	761	7.3

 

Dose (µg/plate)	TA1535
	NA (-)		10% HLI (-)		10% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	9	1.8	9	1.7	12	0.9
100	11	1	6	0.9	8	1.2
333	12	2.8	5	2.0	10	1.2
1000	8	1.2	5	0.9	8	0.3
3333	5P	1.2	5P	2.2	5P	1.2
6666	4P	1
10000			5P	1.5	9P	0.3
Positive control	490	20.9	342	19.1	224	1.5

 

 

Dose (µg/plate)	TA1535
	NA (-)		10% HLI (-)		10% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	9	1.8	10	1.5	11	1.2
100	11	1	9	2.8	9	2.1
333	12	2.8	9	0.9	11	2.3
1000	8	1.2	8	1.7	9	0.6
3333	5P	1.2	9P	1.5	8P	1.7
6666	4P	1
10000			4P	0.6	9P	1.5
Positive control	490	20.9	361	25.9	198	12.5

 

Dose (µg/plate)	TA97
	NA (-)		10% HLI (-)		10% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	129	8.7	136	9.4	157	3.5
100	102	5.0	143	4.7	163	7.8
333	88	8.1	147	5.8	166	6.8
1000	84	3.2	135	6.5	142	2.5
3333	73P	7.8	135P	14.4	121P	4.9
6666	73P	7.8
10000			128P	10.2	134P	13.3
Positive control	1165	125.1	1440	66.5	1223	27.1

                                                                          

 

Dose (µg/plate)	TA97
	NA (-)		30% HLI (-)		30% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	129	8.7	188	6.6	187	10.1
100	102	5.0	181	16.6	187	2.0
333	88	8.1	175	10.7	180	12.4
1000	84	3.2	136	6.4	123	8.9
3333	73P	7.8	113P	5.8	134P	19.2
6666	73P	7.8
10000			112P	6.2	120P	5.3
Positive control	1165	125.1	500	3.2	1251	49.7

 

Dose (µg/plate)	TA98
	NA (-)		10% HLI (-)		10% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	13	2.3	27	1	30	2.8
100	13	0	30	2.7	32	0.9
333	12	2.3	22	2.1	30	1.5
1000	12	2	20	2.7	29	1.2
3333	13P	3.5	19P	1.2	25P	1.7
6666	8P	0.6
10000			22P	3.4	27P	2
Positive control	1115	88.9	1482	54.2	389	17.5

 

Dose (µg/plate)	TA98
	NA (-)		30% HLI (-)		30% RLI (-)
	Mean	SEM	Mean	SEM	Mean	SEM
0.000	13	2.3	19	2.1	19	0.9
100	13	0	24	2.6	32	0.6
333	12	2.3	25	4.3	19	1.2
1000	12	2	23	3.2	27	2.5
3333	13P	3.5	27P	3.5	20P	4.1
6666	8P	0.6
10000			18P	1.5	23P	3.5
Positive control	1115	88.9	1492	153.8	483	12.2

 

 

Conclusions:

The endpoint for the genetic toxicity in vitro of N-ethyl-N phenyl benzyl amine (92-59-1) was considered to be negative in the presence and absence of metabolic activator in Salmonella typhimurium strains TA97, TA98, TA100, and TA1535 by Preincubation method.

Executive summary:

In vitro genetic toxicity test for N-ethyl-N phenyl benzyl amine (92-59-1) was performed on Salmonella typhimurium strains TA97, TA98, TA100, and TA1535 by Preincubation method. The test chemical (0.05 ml), Salmonella culture (0.10 ml), and S-9 mix or buffer (0.50 ml) was incubated at 37°C, without shaking, for 20 min. After that histidine-independent (his⁺) colonies were counted on plates by machine unless there was any precipitate was present which interfered with the count, or the color of the test chemical on the plate reduced the contrast between the colonies and the background agar.The test material was exposed at the concentration of 0.000 ,100.000, 333.000, 1000.000,3333.000,6666.000,10000.000 µg/plate. There are some variations in protocol as the first test of a chemical was without activation and with 10% S-9 in the S-9 mix. If a positive result was obtained the test was repeated.If the tests were negative they were repeated without S-9 and with 30% S-9.Concurrent solvent and positive controls were run with each test. After exposure no mutagenic effect were observed.Therefore N-ethyl-N phenyl benzyl amine (92-59-1) was considered to be non mutagenic in Salmonella typhimurium strains TA97, TA98, TA100, and TA1535in the presence and absence of metabolic activation.

Hence it is not likely to be classified as genetic mutant in vitro.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (negative)

Genetic toxicity in vivo

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Genotoxicity In-vitro

Various publications were reviewed to determine the mutagenic nature of N-ethyl-N phenyl benzyl amine (92-59-1). The studies are as mentioned below:

Gene mutation toxicity study was performed by Errol Zeiger et al (Environmental and Molecular Mutagenesis, 1988) to determine the mutagenic nature of N-ethyl-N phenyl benzyl amine (92-59-1) using Salmonella typhimurium strains. In vitro genetic toxicity test for N-ethyl-N phenyl benzyl amine was performed on Salmonella typhimurium strains TA97, TA98, TA100, and TA1535 by Preincubation method. The test chemical (0.05 ml), Salmonella culture (0.10 ml), and S-9 mix or buffer (0.50 ml) was incubated at 37°C, without shaking, for 20 min. After that histidine-independent (his+) colonies were counted on plates by machine unless there was any precipitate was present which interfered with the count, or the colour of the test chemical on the plate reduced the contrast between the colonies and the background agar. The test material was exposed at the concentration of 0.000, 100.000, 333.000, 1000.000, 3333.000, 6666.000, 10000.000 µg/plate. There are some variations in protocol as the first test of a chemical was without activation and with 10% S-9 in the S-9 mix. If a positive result was obtained the test was repeated. If the tests were negative they were repeated without S-9 and with 30% S-9.Concurrent solvent and positive controls were run with each test. After exposure no mutagenic effect were observed. Therefore N-ethyl-N phenyl benzyl amine (92-59-1) was considered to be non mutagenic in Salmonella typhimurium strains TA97, TA98, TA100, and TA1535in the presence and absence of metabolic activation.

Supported by a experimental study conducted by U.S. National Library of Medicine (CCRIS: N-ETHYL-N-PHENYLBENZYLAMINE, database by TOXNET toxicology data network, 2017), Gene mutation toxicity study in vitro was performed to determine the mutagenic nature of N-ethyl-N phenyl benzyl amine. In genetox study N-Ethyl-N-Phenylbenzylamine (92-59-1) was assessed for its possible mutagenic potential. For this purpose In vitro gene mutation study in bacteria was conducted on S. typhimurium strain TA98, TA100, TA 1535, TA 1537, TA 1538and E.coli WP2UVRA by using preincubation method . DMSO was used as a solvent .The test material was used at concentration of 0, 20-5000 µg/Plate in the presence and absence of metabolic activation. No significant mutagenic effects were observed for N-Ethyl-N-Phenylbenzylaminein the presence and absence of metabolic activator. Therefore N-Ethyl-N-Phenylbenzylamine (92-59-1) was considered to be non mutagenic with and without metabolic activator in S. typhimurium strain TA98, TA100, TA 1535, TA 1537, TA 1538and E.coli WP2UVRA by Ames test. Therefore it is not likely to be classifying as gene mutant in vitro.

It is further supported by prediction data on N-benzyl-N-ethylaniline. Gene mutation toxicity was predicted for N-benzyl-N-ethylaniline (92-59-1) using the battery approach from Danish QSAR database (2017). The study assumed the use of Salmonella typhimurium bacteria in the Ames test. The end point for gene mutation has been modeled in the Danish QSAR using the three software systems Leadscope, CASE Ultra and SciQSAR. Based on predictions from these three systems, a fourth and overall battery prediction is made. The battery prediction is made using the so called Battery algorithm. With the battery approach it is in many cases possible to reduce “noise” from the individual model estimates and thereby improve accuracy and/or broaden the applicability domain.Gene mutation toxicity study as predicted by Danish QSAR for N-benzyl-N-ethylaniline (92-59-1) is negative and hence the chemical is predicted to not classify as a gene mutant in vitro

 

Based on the data and prediction, available for the target chemical, N-benzyl-N-ethylaniline (92-59-1) does not induce gene mutation in vitro. Hence the test chemical is not likely to classify as a gene mutant in vitro.

Justification for classification or non-classification

Thus based on the above annotation and CLP criteria , the target chemical, N-benzyl-N-ethylaniline (92-59-1) is not likely to classify as a gene mutant in vitro.