A mixture of: 2,2',2'',2'''-(ethylenedinitrilotetrakis-N,N-di(C16)alkylacetamide; 2,2',2'',2'''-(ethylenedinitrilotetrakis-N,N-di(C18)alkylacetamide

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:

guideline study with acceptable restrictions

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1991

Report date:

1991

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 471 (Bacterial Reverse Mutation Assay)

Deviations:

yes

Remarks:

strain to detect crosslinking/oxidizing agents was not included

GLP compliance:

yes (incl. QA statement)

Type of assay:

bacterial reverse mutation assay

Test material

Test material information

Constituent 1

Reference substance name:

A mixture of: 2,2',2'',2'''-(ethylenedinitrilotetrakis-N,N-di(C16)alkylacetamide; 2,2',2'',2'''-(ethylenedinitrilotetrakis-N,N-di(C18)alkylacetamide

EC Number:

406-640-0

EC Name:

A mixture of: 2,2',2'',2'''-(ethylenedinitrilotetrakis-N,N-di(C16)alkylacetamide; 2,2',2'',2'''-(ethylenedinitrilotetrakis-N,N-di(C18)alkylacetamide

Cas Number:

136920-07-5

Molecular formula:

Not applicable. Average Molecular Weight: Example Tetra-amide C150H300N6O4 = 2252 g/mol Average Molecular Weight : Based on the average molecular weight of the di-tallow amine starting material (mwt = 509 g/mol) = 2256 g/mol

IUPAC Name:

2-[(2-{bis[(dioctadecylcarbamoyl)methyl]amino}ethyl)[(dioctadecylcarbamoyl)methyl]amino]-N,N-dioctadecylacetamide

Details on test material:

Batch No.: (T 75492/ST 1414/90) Partie 1
Manufacturing/Sampling Date: 10.10.1990
Appearance: Light beige, solid; liquid at 80°C
Storage: 4-6°C

Method

Target gene:

his-

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:

S-9 mix from Aroclor 1254 induced rat livers

Test concentrations with justification for top dose:

20 µg - 5000 µg/plate

Vehicle / solvent:

Tetrahydrofurane (THF)

Controls

Untreated negative controls:

yes

Negative solvent / vehicle controls:

yes

True negative controls:

no

Positive controls:

yes

Positive control substance:

other: with S-9 mix 10 µg 2-aminoanthracene; without S-9 mix 5 µg N-methyl-N -nitro-N-nitrosoguanidine, 10 µg 4-nitro-o-phenylendiamine, 100 µg 9-aminoacridine chloride monohydrate

Details on test system and experimental conditions:

Standard plate test
Test tubes containing 2 ml soft agar kept in a water bath at 45°C, and remaining components added in the following order:
0 .1 ml test solution or vehicle
0 .1 ml bacterial suspension
0 .5 ml S-9 mix (in tests with metabolic activation) or 0 .5 ml phosphate buffer (in tests without metabolic activation).
After mixing, the samples are poured onto Vogel-Bonner agar plates.

Preincubation assay
0.1 ml test solution or vehicle, 0.1 ml bacterial suspension and 0 .5 ml S-9 mix are incubated at 37°C for the duration of 20 minutes. Subsequently, 2 ml of soft agar is added and, after mixing, the samples are poured onto the Vogel-Bonner agar plates.

Standard plate test and Preincubation assay
In each experiment 3 test plates per dose or per control used; after incubation at 37°C for 48 hours in the dark, the bacterial colonies (his+ revertants) are counted. The titer was determined and in regularly measurements the strain characteristics were checked. Sterility control was performed.

Evaluation criteria:

In general, a substance to be characterized as positive in the Ames test has to fulfil the following requirements: doubling of the spontaneous mutation rate (control), dose-response relationship, reproducibility of the results.

Statistics:

none

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 nor precipitates, but tested up to recommended limit concentrations

Vehicle controls validity:

valid

Untreated negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

Solubility: Incomplete solubility of the test substance in tetrahydrofurane (THF) from about 2500 µg/plate onward.

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 (20 - 5000 µg/plate)
Strain	Metabolic activation system	mean revertants in Controls	maximum revertant factor	dose dependency	Assessment
TA 98	no	27	1.0	no	negative
	yes	36	1.0	no	negative
TA 100	no	98	1.1	no	negative
	yes	130	1.2	no	negative
TA 1535	no	16	1.0	no	negative
	yes	19	0.9	no	negative
TA 1537	no	13	1.0	no	negative
	yes	14	1.1	no	negative
Preincubation test (20 - 5000 µg/plate)
Strain	Metabolic activation system	mean revertants in Controls	maximum revertant factor	dose dependency	Assessment
TA 98	no	18	1.3	no	negative
	yes	30	1.3	no	negative
TA 100	no	98	1.2	no	negative
	yes	95	1.4	no	negative
TA 1535	no	15	1.2	no	negative
	yes	13	1.3	no	negative
TA 1537	no	10	1.2	no	negative
	yes	14	1.0	no	negative