Amines, C20-22-alkyldimethyl

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:

2010-02-08/2010-03-15

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: According to the OECD Guideline and it is GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Report date:

2010

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

GLP compliance:

yes

Type of assay:

in vitro mammalian cell micronucleus test

Test material

Test material information

Constituent 1

Reference substance name:

Amines, C20-22-alkyldimethyl

EC Number:

296-866-4

EC Name:

Amines, C20-22-alkyldimethyl

Cas Number:

93164-85-3

Molecular formula:

CnH2nN(CH3)2 n: from 18 to 22

IUPAC Name:

Amines, C20-22-alkyldimethyl

Details on test material:

- Physical state: white solid
- Storage: at room temperature in the dark under nitrogen
- Stability: stable

Method

Target gene:

Histidine

Species / strain

Species / strain 1

Species / strain / cell type:

S. typhimurium TA 1535

Additional strain / cell type characteristics:

other: sodium azide

Species / strain 2

Species / strain / cell type:

S. typhimurium TA 1537

Additional strain / cell type characteristics:

other: 9-aminoacridine

Species / strain 3

Species / strain / cell type:

S. typhimurium TA 98

Additional strain / cell type characteristics:

other: 2-nitrofluorene

Species / strain 4

Species / strain / cell type:

S. typhimurium TA 100

Additional strain / cell type characteristics:

other: methylmethanesulfonate

Species / strain 5

Species / strain / cell type:

other: WP2uvrA

Additional strain / cell type characteristics:

other: 4-nitroquinoline N-oxide

Metabolic activation:

with and without

Metabolic activation system:

S9

Test concentrations with justification for top dose:

The substance was tested in the tester strains TA100 and WP2uvrA with concentrations of 3, 10, 33, 100, 333, 1000, 3330 and 5000 µg/plate in the presence of S9-mix.

Vehicle / solvent:

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

Controls

Untreated negative controls:

no

Negative solvent / vehicle controls:

yes

Positive controls:

yes

Positive control substance:

other: 2-aminoanthracene

Details on test system and experimental conditions:

At least five different doses of the test substance were tested in triplicate in each strain. In the first experiment was tested both in the absence and presence of 5% (v/v) S9-mix in tester strains TA1535, TA1537, and TA98. In an independent repeat of the assay with additional parameters, the test susbtance was tested both in the absence and presence of 10% (v/v) S9-mix in all tester strains. An additonal third experiment with four dose levels was performed with the strains TA13535 and TA1537 in the absence and presence of S9-mix.
The negative control (vehicle) and relevant positive controls were concurrently tested in each strain in the presence and absence of S9-mix.

Results and discussion

Test results

Test results 1

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:

not valid

Positive controls validity:

valid

Test results 2

Species / strain:

other: WP2uvrA

Metabolic activation:

with and without

Genotoxicity:

negative

Cytotoxicity / choice of top concentrations:

no cytotoxicity

Vehicle controls validity:

not 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

Mean number of revertant colonies/3 replicate plates (±S.D.) with different strains of Salmonella typhimurium and one Escherichia coli strain

Dose (µg/plate)	TA1535	TA1537	TA98	TA100	WP2uvrA
without S9 -mix
positive control	834 ± 17	256 ± 72	999 ± 60	861 ± 9	925 ± 25
solvent control	10 ± 4	3 ± 1	19 ±3	93 ± 9	24 ± 6
3				83 ± 8	19 ± 3
10				87 ± 3	15 ± 5
33	10 ± 2	3 ± 2	21 ± 3	78 ± 3	20 ± 7
100	8 ± 3	3 ± 1	17 ± 3	88 ± 8	16 ± 3
333	11 ± 6	4 ± 1	22 ± 3	85 ± 5	19 ± 10
1000	14 ± 6	10 ± 2	15 ± 4	94 ± 1	17 ± 2
3330 SP	13 ± 7	13 ± 9	21 ± 2	53 ± 5	21 ± 6
5000 SP				28 ± 14 s	24 ± 2
with S9 -mix (1)
positive control	209 ± 33	305 ± 80	1420 ± 65	1187 ± 87	329 ± 8
solvent control	7 ± 3	4 ± 1	29 ± 4	77 ± 19	18 ± 6
3				62 ± 7	17 ± 4
10				68 ± 4	14 ± 3
33	6 ± 2	4 ± 1	20 ± 2	70 ± 15	17 ± 4
100	5 ± 1	4 ± 2	32 ± 8	70 ± 3	16 ± 3
333	7 ± 3	3 ± 1	16 ± 2	86 ± 14	18 ± 4
1000	9 ± 5	6 ± 3	29 ± 5	94 ± 5	16 ± 8
3330 SP	11 ± 1	16 ± 2	32 ± 4	100 ± 48	18 ± 8
5000 SP				55 ± 18s	12 ± 2

Solvent control: 0.1 mL ethanol

1 The S9 -mix contained 5% (v/v) S9 fraction

s Bacterial background lawn slightly reduced

sp Slight Precipitate

Mean number of revertant colonies/3 replicate plates (±S.D.) with different strains of Salmonella typhimurium and one Escherichia coli strain

Dose (µg/plate)	TA1535	TA1537	TA98	TA100	WP2uvrA
without S9 -mix
positive control	785 ± 42	284 ±34	946 ±60	1280 ±8	1031 ±29
solvent control	7 ± 1	6 ±1	20 ±1	76 ±10	19 ±4
33	4 ±2	4 ±2	19 ±2	84 ±10	13 ±2
100	7 ±5	6 ±2	20 ±3	80 ±5	19 ±2
333	7 ±3	5 ±4	32 ±5	84 ±4	13 ±3
1000	7 ±1	14 ±7	41 ±27	90 ±3	17 ±3
3330 SP	22 ±5	32 ±2	31 ±6	112 ±5	25 ±3
With S9 -mix (1)
positive control	149 ±5	367 ±40	537 ±10	977 ±37	296 ±16
solvent control	4 ±2	5 ±2	21 ±3	69 ±6	12 ±4
33	7 ±2	4 ±1	21 ±1	62 ±4	9 ±5
100	3 ±0	3 ±0	20 ±1	59 ±2	11 ±4
333	7 ±3	5 ±2	15 ±3	71 ±3	14 ±2
1000	8 ±6	12 ±4	30 ±11	71 ±3	19 ±9
3330 SP	21 ±6	36 ±6	33 ±10	84 ±7	30 ±5

Solvent control: 0.1 mL ethanol

1: The S9 -mix contained 10% (v/v) S9 fraction

SP Slight Precipitate

Mean number of revertant colonies/3 replicate plates (±S.D.) with two Salmonella typhimurium strains

Dose (µg/plate)	TA1535	TA1537
without S9 -mix
positive control	894 ± 39	298 ± 12
solvent control	13 ± 7	4 ± 2
333	16 ± 4	5 ± 3
1000	14 ± 3	4 ± 1
1666 SP	22 ± 4	11 ± 4
3330 SP	21 ± 2	3 ± 2
with S9 -mix
positive control	143 ± 5	341 ± 94
solvent control	11 ± 2	6 ± 3
333	6 ± 1	4 ± 1
1000	14 ± 3	7 ± 2
1666 SP	16 ± 3	5 ± 1
3330 SP	18 ± 4	3 ± 1

Solvent control: 0.1 mL ethanol

1 The S9 -mix contained 10% (v/v) S9 fraction

SP Slight Precipitate

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information):
negative

Based on the results of this study it is concluded that the substance is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.

Executive summary:

The study procedures described in this report were based on the Guideline OECD 471 and B.13/14. The substance was tested in the Salmonella typhimurium reverse mutation assay with four histidine-requiring strains of Salmonella typhimurium (TA1535, TA1537, TA98 and TA100) and in the Escherichia coli reverse mutation assay with a tryptophan-requiring strain of Escherichia coli. The test was performed in two independent experiments in the presence and absence of S9 -mix. To obtain more information about the possible mutagenicity of the susbtance, an additional experiment was performed with the tester strains TA1535 and TA1537 in the absence and presence of 10% (v/v) S9 -mix.

Based on the results of this study it is concluded that the substance is not mutagenic in the Salmonella typhimurium reverse mutation assay and in the Escherichia coli reverse mutation assay.