Triethylammonium bromide

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

Study period:

October 2013 - November 2013

Reliability:

1 (reliable without restriction)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Report date:

2013

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)

Deviations:

no

Principles of method if other than guideline:

None as known.

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

Triethylammonium bromide

EC Number:

211-263-8

EC Name:

Triethylammonium bromide

Cas Number:

636-70-4

Molecular formula:

C6H15N.BrH

IUPAC Name:

N,N-diethylethanaminium bromide

Test material form:

solid: crystalline

Details on test material:

Name triethylammonium bromide
Batch no. 32541
Appearance white crystals
Composition triethylammonium bromide, water
CAS No. 636-70-4
EINECS-No. 211-263-8
Molecular formula C6H16BrN
Molecular weight 182.10 g/mol
Purity 98.2 % (argentometry)
Homogeneity homogeneous
Production date 11. Sep. 2013
Expiry date 10. Sep. 2028
Storage room temp. (20 ± 5°C); kept away from light

Method

Species / strain

Species / strain / cell type:

other: Salmonella typhimurium (TA 97a, TA 98, TA 100, TA 102 and TA 1535)

Metabolic activation:

with and without

Metabolic activation system:

S9 (produced from the livers of male Sprague-Dawley rats which were treated with 500 mg Aroclor 1254/kg body weight intraperitoneally)

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: Deionised water
- Justification for choice of solvent/vehicle: because the test item was sufficiently soluble, and this solvent doesn’t have any effects on the viability of the bacteria or the number of spontaneous revertants.

Controls

Untreated negative controls:

yes

Remarks:

Deionised water

Negative solvent / vehicle controls:

yes

Remarks:

Deionised water

True negative controls:

yes

Remarks:

DMSO

Positive controls:

yes

Positive control substance:

sodium azide

benzo(a)pyrene

other: 4-Nitro-1,2-phenylene diamine; 2-Amino-anthracene

Results and discussion

Test results

Species / strain:

other: Salmonella typhimurium (TA 97a, TA 98, TA 100, TA 102 and TA 1535)

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

First Experiment

Confirmation of the Criteria and Validity

The treatments for the sterility control and the determination of the titre didn’t show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range of the test laboratory. All positive controls (diagnostic mutagenes) showed mutagenic effects with and without metabolic activation.

Solubility and Toxicity

The test item was dissolved in deionised water. A stock solution containing 50.0±0.5 g/L was prepared.

No signs of toxicity towards the tested strains could be observed. The background lawn was visible and the number of revertant colonies was not reduced.

Main Revertants First Experiment

Strain		TA97a		TA98		TA100		TA102		TA1535
Induction		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
H2O	Mean	99	106	15	17	151	135	186	220	21	21
	sd	13.7	16.1	2.2	4.5	3.3	21.6	9.4	9.9	3.3	5.4
DMSO	Mean	128	105	17	18	152	122	186	217	19	17
	sd	22.9	10.4	2.2	2.2	7.4	10.1	9.8	17.4	4.3	7.8
Positive Controls*	Mean	590	652	118	95	689	630	540	449	108	125
	sd	97	39	10	10	31	89	111	20	14	12
	f(I)	4.61	6.21	6.94	5.28	4.56	5.16	2.90	2.07	5.14	7.35
5007 µg/pl.	Mean	121	119	20	18	135	114	181	203	22	21
	sd	7	8	2	3	14	24	14	23	3	2
	f(I)	1.22	1.12	1.33	1.06	0.89	0.84	0.97	0.92	1.05	1.00
1502 µg/pl.	Mean	118	103	17	18	151	132	194	216	22	20
	sd	21	6	2	6	25	10	37	14	5	4
	f(I)	1.19	0.97	1.13	1.06	1.00	0.98	1.04	0.98	1.05	0.95
501 µg/pl.	Mean	114	111	15	19	126	132	192	198	25	18
	sd	9	27	3	2	10	7	7	22	6	2
	f(I)	1.15	1.05	1.00	1.12	0.83	0.98	1.03	0.90	1.19	0.86
150 µg/pl.	Mean	105	113	19	17	141	122	194	227	22	20
	sd	15	19	3	1	3	26	15	19	6	2
	f(I)	1.06	1.07	1.27	1.00	0.93	0.90	1.04	1.03	1.05	0.95
50 µg/pl.	Mean	122	119	18	21	154	133	208	234	24	20
	sd	9	3	2	4	11	39	14	7	2	3
	f(I)	1.23	1.12	1.20	1.24	1.02	0.99	1.12	1.06	1.14	0.95

f(I) = increase factor

* Different positive controls were used

Mutagenicity

No significant increase of the number of revertant colonies in the treatments with and without metabolic activation could be observed.No concentration-related increase over the tested range was found.

 

Therefore, the test item is stated as not mutagenic under the test conditions.

To verify this result, a second experiment was performed using the pre-incubation method.

Second Experiment

Confirmation of the Criteria and Validity

The treatments for the sterility control and the determination of the titre didn’t show any inconsistencies. The determined values for the spontaneous revertants of the negative controls were in the normal range of the test laboratory. All positive controls showed mutagenic effects with and without metabolic activation.

Solubility and Toxicity

The test item was dissolved in deionised water. A stock solution containing 50.0±0.5 g/L was prepared.

No signs of toxicity towards the tested strains could be observed. The background lawn was visible and the number of revertant colonies was not significantly reduced.

Survey of the Findings

The mean revertant values of the four replicates are presented in the following table.

Mean Revertants Second Experiment

Strain		TA97a		TA98		TA100		TA102		TA1535
Induction		-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9	-S9	+S9
H2O	Mean	100	114	14	10	115	121	207	196	19	19
	sd	6.2	16.6	3.0	0.0	9.5	16.9	17.5	24.1	2.9	1.3
DMSO	Mean	105	113	15	13	118	117	198	205	20	21
	sd	3.6	11.0	1.3	0.0	13.4	12.8	20.8	28.3	4.3	3.0
Positive Controls*	Mean	581	767	109	146	569	957	676	804	155	140
	sd	151	165	4	49	116	253	56	161	23	36
	f(I)	5.53	6.79	7.27	11.23	4.95	8.18	3.41	3.92	8.16	6.67
5023 µg/pl.	Mean	116	131	15	16	123	119	237	198	21	21
	sd	15	27	2	2	17	22	7	14	3	4
	f(I)	1.16	1.15	1.07	1.60	1.07	0.98	1.14	1.01	1.11	1.11
2512 µg/pl.	Mean	103	118	14	15	127	107	226	178	19	17
	sd	7	13	3	2	14	6	31	16	2	2
	f(I)	1.03	1.04	1.00	1.50	1.10	0.88	1.09	0.91	1.00	0.89
1256 µg/pl.	Mean	105	118	16	12	139	129	211	188	24	21
	sd	4	13	2	2	13	20	22	29	2	2
	f(I)	1.05	1.04	1.14	1.20	1.21	1.07	1.02	0.96	1.26	1.11
628 µg/pl.	Mean	106	119	17	13	114	119	222	174	19	21
	sd	11	28	4	1	21	17	25	15	3	2
	f(I)	1.06	1.04	1.21	1.30	0.99	0.98	1.07	0.89	1.00	1.11
314 µg/pl.	Mean	98	107	16	12	129	117	194	207	22	22
	sd	3	13	3	1	8	8	28	22	2	2
	f(I)	0.98	0.94	1.14	1.20	1.12	0.97	0.94	1.06	1.16	1.16

f(I) = increase factor

* Different positive controls were used

Mutagenicity

No significant increase of the number of revertant colonies in the treatments with and without metabolic activation was observed.

No concentration-related increase over the tested range was found.

Therefore, the test item is stated as not mutagenic under the test conditions.

 

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

The test item triethylammonium bromide is considered as “not mutagenic under the conditions of the test”.

Executive summary:

The test item is considered not mutagenic for the reasons given above. The test item did not show any cytotoxicity towards the bacteria.

The confirmation tests of the genotype didn’t show any irregularities. The control of the titre was above the demanded value.

The numbers of revertant colonies of the positive controls were within the range of the historical data (all batch numbers of bacteria) of the laboratory and were definitely increased in comparison with the negative controls, as well as showing mutagenic potential of the diagnostic mutagenes.

Spontaneous revertants were within the normal range in comparison with the historical data (all batch numbers of bacteria) of the LAUS GmbH. For these reasons, the result of the test is considered valid.