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EC number: 217-461-0 | CAS number: 1860-26-0
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- Ecotoxicological Summary
- Aquatic toxicity
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- Short-term toxicity to fish
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Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- comparable to guideline study with acceptable restrictions
- Remarks:
- only four strains used in study, only one positive control used for the assay with metabolic activation
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 1 985
- Report date:
- 1985
Materials and methods
Test guideline
- Qualifier:
- equivalent or similar to guideline
- 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) and in accordance with OECD 471 from 26 May 1983, which was replaced.
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 substance name:
- 2-ethyl-N,N-bis(2-ethylhexyl)hexylamine
- EC Number:
- 217-461-0
- EC Name:
- 2-ethyl-N,N-bis(2-ethylhexyl)hexylamine
- Cas Number:
- 1860-26-0
- Molecular formula:
- C24H51N
- IUPAC Name:
- tris(2-ethylhexyl)amine
- Test material form:
- liquid
- Details on test material:
- - Name of test material (as cited in study report): Tri-2-Aethylhexylamin
- Physical state: liquid
- Analytical purity: approx. 98%
Constituent 1
- Specific details on test material used for the study:
- - 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
- Details on mammalian cell type (if applicable):
- MEDIA USED
- Type and composition of media: nutrient broth solution (8 g Difco bacto nutrient broth + 5 g NaCl/liter)
- temperature: 37°C
- Metabolic activation:
- with and without
- Metabolic activation system:
- Type and composition of metabolic activation system:
- source of S9 : 5 male Sprague-Dawley rats, injected with Aroclor 1254, 5 days before sacrifice
- method of preparation of S9 mix: livers washed, homogenized in three volumes of KCl solution (150 mM), centrifugation at 9,000 g for 10 min, use of supernatent - Test concentrations with justification for top dose:
- 20, 100, 500, 2500 and 5000 µg/plate
- Vehicle / solvent:
- - Ssolvent 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:
- 9-aminoacridine
- 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
- 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
- True negative controls validity:
- not applicable
- 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.
Any other information on results incl. tables
Table 1: Standard plate test 1 (Mean ± SD)
Dose (µg/plate) | TA 1535 | TA 100 | TA 1537 | TA 98 | ||||
-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 | - |
Table 2: Standard plate test 2 (Mean ± SD)
Dose (µg/plate) | TA 1535 | TA 100 | TA 1537 | TA 98 | ||||
-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 | - |
2-AA: 2-aminoanthracene;
MNNG; N-methyl-N-nitro-N-nitrosoguanidine
NPD: 4-nitro-o-phenylendiamine
AAC: 9-aminoacridine chloride monohydrate
Applicant's summary and conclusion
- Conclusions:
- Under the conditions tested the test substance is non mutagenic in the bacterial AMES-test.
The positive control gave the expected values.
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