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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:
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