2-ethyl-N,N-bis(2-ethylhexyl)hexylamine

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:

other: see 'Remark'

Remarks:

Acceptable, well documented report which meets basic scientific principles. Comparable to guideline study with acceptable restrictions (neither Escherichia coli strain WP2 uvrA pKM101a nor S. typhimurium strain TA102 were employed in this study for the detection of oxidising mutagens and cross-linking agents; 2-aminoanthracene used as single positive control for the assays with metabolic activation).

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1985

Report Date:

1985

Materials and methods

Test guideline

Qualifier:

equivalent or similar to

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

no strain used for the detection of oxidising mutagens and cross-linking agents; 2-aminoanthracene used as single positive control for the assays with metabolic activation

Principles of method if other than guideline:

The study was conducted according to the testing protocol published by Ames (1973, 1975).
Ames BN et al. (1973). Proc. Nat. Acad. Sci. USA 70: 2281 -2285
Ames BN et al. (1975). Mut. Res. 31: 347 - 364

GLP compliance:

no

Type of assay:

bacterial reverse mutation assay

Test material

Reference

Name:

Unnamed

Type:

Constituent

Details on test material:

- Name of test material (as cited in study report): Tri-2-ethylhexylamine
- Physical state: liquid
- Analytical purity: approx. 98 %
- Impurities (identity and concentrations): approx. 1.5 % Di-2-ethylhexylamine

Method

Target gene:

Histidine operon

Species / strain

Species / strain / cell type:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

Metabolic activation:

with and without

Metabolic activation system:

S9-mix from Aroclor 1254 treated male Sprague-Dawley rats.

Test concentrations with justification for top dose:

0, 20, 100, 500, 2500 and 5000 µg/plate

Vehicle / solvent:

- Vehicle(s)/solvent(s) used: acetone

Controls

Untreated negative controls:

yes

Remarks:

sterility control

Negative solvent / vehicle controls:

yes

Remarks:

water control

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S-9 mix: all strains 2-aminoanthracene; without S-9 mix: strains TA100, TA1535: N-methyl-N'-nitro-N-nitrosoguanidine; TA98: 4-nitro-o-phenylendiamine; TA1537: 9-aminoacridine

Details on test system and experimental conditions:

METHOD OF APPLICATION: standard plate test

DURATION
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 test plates per test concentration or control; two experiments were carried out independently.

Evaluation criteria:

The test chemical is considered positive in this assay if the following criteria are met:
- A dose-related and reproducible increase in the number of revertant colonies, i.e. about doubling of the spontaneous mutation rate in at least one tester strain either without S-9 mix or after adding a metabolizing system.

A test substance is generally considered non-mutagenic in this test if:
- The number of revertants for all tester strains were within the historical negative control range under all experimental conditions in two experiments carried out independently of each other.

Results and discussion

Test results

Species / strain:

S. typhimurium TA 1535, TA 1537, TA 98 and TA 100

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

Additional information on results:

TEST-SPECIFIC CONFOUNDING FACTORS
- Water solubility: Complete solubility of the test substance in acetone up to the highest dose of 5000 µg/plate.

Remarks on result:

other: all strains/cell types tested

Remarks:

Migrated from field 'Test system'.

Any other information on results incl. tables

Standard plate test 1.:

Dose (µg/plate)

TA1535

TA100

TA1537

TA98

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

0

17±4

14±5

130±6

113±6

6±1

6±1

27±3

37±5

20

14±3

11±1

135±26

142±9

10±2

9±2

21±4

34±7

100

16±5

14±1

127

136±9

10±2

12±5

18±6

34±4

500

17±5

13±1

152±17

139±10

9±2

12±5

22±3

33±6

2500

16±2

16±3

118±20

132±5

11±2

10±1

25±1

28±6

5000

17±4

13±6

124±6

152±8

5±3

10±1

23±3

19±1

2-AA

-

453±76

-

2500±278

-

141±42

-

1423±45

MNNG

2367±29

-

2467±333

-

-

-

-

-

AAC

-

-

-

-

60±16

-

-

-

NPD

-

-

-

-

-

-

563±140

-

Mean ± SD

Standard plate test 2.:

Dose (µg/plate)

TA1535

TA100

TA1537

TA98

-S9

+S9

-S9

+S9

-S9

+S9

-S9

+S9

0

15±3

14±4

101±6

108±4

7±2

7±1

27±4

41±6

20

14±4

12±2

94±12

128±10

7±1

6±2

28±6

31±3

100

14±4

13±1

107±7

113±10

5±1

8±2

21±9

31±3

500

10±1

16±5

96±11

107±3

4±1

8±2

25±5

31±3

2500

11±1

16±2

87±8

110±15

5±1

4±1

27±5

30±4

5000

11±1

13±5

83±4

115±3

4±2

3±1

29±5

37±9

2-AA

-

319±20

-

2017±206

-

211±15

-

1533±166

MNNG

1640±284

-

1910±62

-

-

-

-

-

AAC

-

-

-

-

632±35

-

-

-

NPD

-

-

-

-

-

-

873±44

-

Mean ± SD

2-AA: 2-aminoanthracene;

MNNG; N-methyl-N-nitro-N-nitrosoguanidine

NPD: 4-nitro-o-phenylendiamine

AAC: 9-aminoacridine chloride monohydrate

Under the conditions tested Tri-2-ethylhexylamine is non mutagenic in the bacterial AMES-test.

The positive control gave the expected values.

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative