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Toxicological information

Genetic toxicity: in vitro

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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:
OECD guideline study conducted in compliance with GLP regulations, with acceptable restrictions (neither E. coli strain WP2 uvrA nor S. typhimurium strain TA102 were employed in this study for the detection of oxidising mutagens and cross-linking agents)

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1999
Report date:
1999

Materials and methods

Test guideline
Qualifier:
according to guideline
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
(adopted 1983)
Deviations:
yes
Remarks:
(neither E. 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 sole indicator of the efficacy of the S9 mix)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Constituent 1
Chemical structure
Reference substance name:
Amines, tri-C8-10-alkyl
EC Number:
272-347-8
EC Name:
Amines, tri-C8-10-alkyl
Cas Number:
68814-95-9
Molecular formula:
Unspecified
IUPAC Name:
N,N-dinonylnonan-1-amine
Test material form:
other: liquid
Details on test material:
- Name of test material (as cited in study report): Alamine 336
- Physical state: liquid, colourless
- Analytical purity: 100 % (active substance)
- Lot/batch No.: 5J167
- Expiration date of the lot/batch: February 2000
- Stability under test conditions: stable for 12 months
- Storage condition of test material: at room temperature
- SAT-No.: 990409

Method

Target gene:
his operon
Species / strain
Species / strain / cell type:
S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Metabolic activation system:
Phenobarbital/B-Naphthoflavone induced rat liver S-9 mix
Test concentrations with justification for top dose:
1st and 2nd assay: 8, 40, 200, 1000 and 5000 µg/plate
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: Tween 80/bidest. water
- Justification for choice of solvent/vehicle: The suspension medium was chosen according to the solubility properties tested preliminary before start of the study.
Controls
Untreated negative controls:
yes
Remarks:
untreated cell suspension
Negative solvent / vehicle controls:
yes
Remarks:
Tween 80/bidest. water
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
sodium azide
other: 4-nitro-o-phenylenediamine, 2-aminoanthracene
Details on test system and experimental conditions:
METHOD OF APPLICATION: in agar (plate incorporation)

DURATION
- Preincubation period: 12 hours

NUMBER OF REPLICATIONS: triplicate plates

DETERMINATION OF CYTOTOXICITY
- Method: background lawn

DETAILS ON CONTROLS:
- Negative controls: suspension medium Tween 80/bidist. water and the untreated fresh cell suspensions
- Positive controls:
- Without metabolic activation:
sodium azide (2 µg/plate) for TA 100 and TA 1535;
9-aminoacridine (80 µg/plate) for TA 1537
4-nitro-o-phenylendiamine (40 µg/plate) for TA 98 and TA 1538
- With metabolic activation:
2-aminoanthracene in all strains (2.5 µg/plate for TA 1535, TA 1537; 5.0 µg/plate for TA 100, TA 98, TA 1538)
- Sterility controls:
The test batches containing S9-mix were each controlled for sterility by adding 0.5 mL of S9-mix fraction to untreated agar plates.

Results and discussion

Test results
Species / strain:
S. typhimurium, other: TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
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

RESULTS:

 

Experiment 1

Treatment groups

[µg/plate]

Strains

 

 

TA 100

TA 1535

TA 1537

TA 1538

TA 98

 

 

-

SD

+

SD

-

SD

+

SD

-

SD

+

SD

-

SD

+

SD

-

SD

+

SD

medium

-

108

6

125

15

9

3

18

12

16

5

10

0

10

4

11

6

19

8

32

2

solvent

-

129

15

117

14

10

1

13

1

17

5

11

3

8

1

12

7

22

6

25

4

test substance

8

121

18

137

17

10

2

11

3

14

3

20

2

10

3

9

6

29

8

22

3

40

121

15

134

16

14

5

14

4

9

2

13

4

15

5

11

2

24

3

27

2

200

115

12

113

17

7

2

11

3

14

1

4

3

13

3

15

3

23

8

31

5

1000

113

11

98

9

9

1

11

6

13

3

12

3

8

3

11

3

20

4

29

5

5000

106

8

87

16

11

4

7

4

5

1

5

2

11

5

14

1

14

3

22

7

positive

controls

A

736

40

302

6

542

8

222

13

906

59

463

56

2421

54

1784

75

1718

100

1381

166

B

148

13

2990

18

11

1

194

35

15

6

181

15

17

3

1992

122

19

3

2125

110

Experiment 2

Treatment groups

[µg/plate]

Strains

 

 

TA 100

TA 1535

TA 1537

TA 1538

TA 98

 

 

-

SD

+

SD

-

SD

+

SD

-

SD

+

SD

-

SD

+

SD

-

SD

+

SD

medium

-

103

12

116

15

8

5

10

2

14

5

20

7

13

7

18

8

23

3

20

6

solvent

-

111

15

96

16

24

2

9

3

10

2

12

1

16

7

12

2

26

4

24

2

test substance

8

115

6

117

15

14

7

16

1

10

1

15

5

10

3

17

6

23

0

23

6

40

111

3

125

24

12

4

15

4

14

3

10

4

13

1

14

3

23

2

28

2

200

106

11

119

23

15

2

13

5

7

1

8

3

13

4

13

3

21

2

30

5

1000

81

11

82

3

12

5

10

1

7

2

9

5

12

4

12

3

13

2

23

2

5000

82

11

86

9

152

2

12

6

8

3

10

5

8

2

11

1

13

1

25

2

positive

controls

A

764

19

165

22

615

21

76

9

409

120

112

28

2387

235

1227

297

1587

21

872

96

B

118

9

943

91

27

9

115

25

18

4

78

8

16

1

951

123

27

6

727

75

-: without metabolic activation

+: with metabolic activation

SD: standard deviation

Positive controls:         A:        TA 1535, TA 100:                     sodium azide

                                           TA 1537:                                9-aminoacridine

                                           TA 1538, TA 98:            4-nitro-o-phenylenediamine

                                B :       TA 1535, TA 1537:                    2-aminoanthracene

                                           TA 100, TA 1538, TA 98:            2-aminoanthracene

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

Amines, tri-C8-10-alkyl is considered not to be mutagenic in this bacterial mutagenicity test in vitro.
Executive summary:

The test item with the batch number 5J167 was tested in the Salmonella typhimurium Reverse Mutation Assay for the induction of reverse mutations in a bacterial test system. The assay was performed with the Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537 and TA 1538 in two independent experiments, both with and without metabolic activation by S9-mix according to OECD 471 (BASF SE, 1999, 9901542).

Solutions of the test substance were prepared in Tween 80/bidist. water and diluted with Tween 80/bidist. water just before use. The following concentrations were tested:

1st and 2nd test: 8, 40, 200, 1000 and 5000 µg/plate

The direct plate incorporation assay was utilized. The following results were obtained:

Toxic effects were not noted. No enhanced revertant rates compared to concurrent negative controls were observed in all tested strains, neither in the presence nor in the absence of metabolic activation.

Therefore the test substance did not induce reverse mutations in the tested strains of Salmonella typhimurium in this bacterial mutagenicity test.

In conclusion, it can be stated that in the study described and under the experimental conditions reported, the test substance did not induce gene mutations in Salmonella typhimurium strains.

Subsequently, Amines, tri-C8-10-alkyl is considered not to be mutagenic in this

bacterial mutagenicity test in vitro.

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