Registration Dossier
Registration Dossier
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 249-044-4 | CAS number: 28472-97-1
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- migrated information: read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- GLP - Guideline study, tested with the source substance bis(2-ethylhexyl)nonanedioate (CAS 103-24-2). In accordance to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance.
Data source
Referenceopen allclose all
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 004
- Report date:
- 2004
- Reference Type:
- publication
- Title:
- Reverse Mutation Test of Bis(2-ethylhexyl) azelate in Bacteria
- Author:
- Miwa, Y.
- Year:
- 2 004
- Bibliographic source:
- Toxicity Testing Reports of Environmental Chemicals, 11, 287-320
- Reference Type:
- secondary source
- Title:
- Bis(2-ethylhexyl) azelate
- Author:
- OECD
- Year:
- 2 006
- Bibliographic source:
- SIDS Initial Assessment Report for SIAM 22, Paris, France, 18-21 April, 2006
- Reference Type:
- secondary source
- Title:
- Diesters Category of the Aliphatic Esters Chemicals (Test Plan and Robust Summaries for Substances in the HPV Test Plan)
- Author:
- US-EPA (American Chemistry Council's Aliphatic Esters Panel)
- Year:
- 2 010
- Bibliographic source:
- High Production Volume (HPV) Chemical Challenge Program (201-16837A and 201-16837B)
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
Test material
- Reference substance name:
- 103-24-2 (analytical purity 77.2%)
- IUPAC Name:
- 103-24-2 (analytical purity 77.2%)
- Details on test material:
- - Name of test material (as cited in study report): Bis(2-ethylhexyl)nonanedioate
- Analytical purity: 77.2%
- Impurities (identity and concentrations):
Bis(2-ethylhexyl)glutarate 2.2%,
Bis(2-ethylhexyl)adipate 2.4%,
Bis(2-ethylhexyl)pimelate 2.8%,
Bis(2-ethylhexyl)suberate 3.8%,
Bis(2-ethylhexyl)sebacate 3.3%,
Bis(2-ethylhexyl) 1-,9-nonamethylenedicarboxylate 5.3%,
Bis(2-ethylhexyl)1-,10-decamethylenedicarboxylate 0.6%,
Bis(2-ethylhexyl)1-,11-undecamethylenedicarboxylate 0.3%
- Lot/batch No.: N-31101
Constituent 1
Method
- Target gene:
- His operon (S. typhimurium)
Trp operon (E. coli)
Species / strainopen allclose all
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Species / strain / cell type:
- E. coli WP2 uvr A
- Metabolic activation:
- with and without
- Metabolic activation system:
- cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with phenobarbital and 5,6-benzoflavone.
- Test concentrations with justification for top dose:
- Range finding test:
1.22, 4.88, 19.5, 78.1, 312.5, 625, 1250 and 5000 µg/plate with and without metabolic activation
Main test:
312.5, 625, 1250, 2500 and 5000 µg/plate with and without metabolic activation - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
Controls
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: -S9: 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide (AF2; 0.01 µg/plate: TA100, WP2uvrA; 0.1 µg/plate: TA98); sodium azide (SA; 0.5 µg/plate: TA1535); 9-aminoacridine (9AA; 80 µg/plate: TA1537); +S9: 2-aminoanthracene (2AA; up to 10 µg/plate all strains)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation)
DURATION
- Exposure duration: 48 h
NUMBER OF REPLICATIONS: triplicates each in two independent experiments
DETERMINATION OF CYTOTOXICITY
- Method: relative total growth - Evaluation criteria:
- If the number of colonies with revertants was as double as that of control group and dose response was observed, the interpretation of the result was positive.
- Statistics:
- Mean values and standard deviation were calculated.
Results and discussion
Test resultsopen allclose all
- 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
- Species / strain:
- E. coli WP2 uvr A
- 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:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: There were colorless clear fine oil drop-like precipitations and oil membrane-like precipitations on the surface of agar in any test concentration.
RANGE-FINDING: No cytotoxicity was observed in a range finding study with doses up to 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
Table 1. Test results of experiment 1 (plate incorporation)
With or without S9-Mix |
Test substance concentration |
Mean number of revertant colonies per plate |
||||
(μg/plate) |
(average of 3 plates ± Standard deviation) |
|||||
|
Base-pair substitution type |
cross-linking type |
Frameshift type |
|||
|
TA 100 |
TA1535 |
WP2 uvrA |
TA98 |
TA1537 |
|
– |
0 (aqua dest.) |
102 ± 8.1 |
9 ± 2.6 |
35 ± 2.6 |
20 ± 3.5 |
5 ± 0.6 |
– |
0 (vehicle) |
110 ± 14 |
13 ± 6.2 |
27 ± 2.1 |
29 ± 4.2 |
7 ± 2.3 |
– |
312.5 |
100 ± 9.3P |
8 ± 2.1P |
25 ± 7.0P |
23 ± 0.6P |
13 ± 3.1P |
– |
625 |
106 ± 2.6P |
11 ± 2.6P |
31 ± 6.5P |
23 ± 4.2P |
11 ± 0.6P |
– |
1250 |
112 ± 12.1P |
10 ± 1.2P |
26 ± 5.5P |
29 ± 2.1P |
6 ± 0.6P |
– |
2500 |
94 ± 2.5P |
9 ± 2.5P |
29 ± 2.6P |
27 ± 4.0P |
10 ± 3.5P |
– |
5000 |
92 ± 11.5P |
7 ± 1.0P |
24 ± 1.5P |
23 ± 5.0P |
4 ± 0.6P |
Positive controls, |
Name |
AF2 |
SA |
AF2 |
AF2 |
9AA |
Concentrations (μg/plate) |
0.01 |
0.5 |
0.01 |
0.1 |
80 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
548 ± 7.8 |
525 ± 7.1 |
132 ± 6.5 |
421 ± 11.2 |
334 ± 87.7 |
|
+ |
0 (medium) |
106 ± 12.1 |
11 ± 0.6 |
24 ± 3.5 |
30 ± 3.0 |
13 ± 1.0 |
+ |
0 (vehicle) |
105 ± 12 |
10 ± 4.0 |
33 ± 7.2 |
27 ± 3.5 |
16 ± 3.8 |
+ |
312.5 |
103 ± 8.1P |
7 ± 4.4P |
30 ± 6.1P |
25 ± 4.6P |
13 ± 1.7P |
+ |
625 |
96 ± 8.1P |
7 ± 2.6P |
31 ± 4.6P |
31 ± 8.0P |
11 ± 1.5P |
+ |
1250 |
97 ± 15.6P |
9 ± 2.6P |
31 ± 5.0P |
31 ± 6.4P |
14 ± 1.5P |
+ |
2500 |
96 ± 9.7P |
6 ± 3.2P |
33 ± 2.6P |
25 ± 7.5P |
11 ± 2.0P |
+ |
5000 |
90 ± 3.2P |
7 ± 4.2P |
32 ± 5.8P |
32 ± 5.5P |
12 ± 2.9P |
Positive controls, |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentrations (μg/plate) |
1 |
2 |
10 |
0.5 |
2 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
1036 ± 46.9 |
327± 18.4 |
969 ± 142.3 |
437 ± 30.3 |
181 ± 29.5 |
|
AF2 = 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide
9AA = 9-aminoacridine
2AA = 2-aminoanthracene
SA = sodium azide
P = Precipitate
Table 2. Test results of experiment 2 (plate incorporation)
With or without S9-Mix |
Test substance concentration |
Mean number of revertant colonies per plate |
||||
(μg/plate) |
(average of 3 plates ± Standard deviation) |
|||||
|
Base-pair substitution type |
cross-linking type |
Frameshift type |
|||
TA 100 |
TA1535 |
WP2 uvrA |
TA98 |
TA1537 |
||
– |
0 (aqua dest.) |
132 ± 15.5 |
8 ± 1.0 |
39 ± 2.5 |
23 ± 8.1 |
9 ± 5.5 |
– |
0 (vehicle) |
131 ± 14.5 |
9 ± 3.1 |
41 ± 6.1 |
31 ± 6.9 |
9 ± 3.0 |
– |
312.5 |
129 ± 17.1P |
7 ± 2.1P |
38 ± 7.0P |
27 ± 4.2P |
10 ± 1.5P |
– |
625 |
124 ± 5.2P |
7 ± 1.2P |
33 ± 7.8P |
30 ± 11P |
14 ± 5.2P |
– |
1250 |
127 ± 10.4P |
8 ± 1.5P |
34 ± 4.0P |
30 ± 2.1P |
13 ± 1.0P |
– |
2500 |
128 ± 16.8P |
8 ± 3.8P |
38 ± 3.2P |
25 ± 1.0P |
13 ± 3.6P |
– |
5000 |
124 ± 4.0P |
10 ± 2.5P |
38 ± 0.0P |
27 ± 4.0P |
10 ± 2.5P |
Positive controls, |
Name |
AF2 |
SA |
AF2 |
AF2 |
9AA |
Concentrations (μg/plate) |
0.01 |
0.5 |
0.01 |
0.1 |
80 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
569 ± 27.5 |
589 ± 36.5 |
137 ± 17.6 |
446 ± 19.1 |
411 ± 100.1 |
|
+ |
0 (medium) |
137 ± 16.1 |
7 ± 2.5 |
33 ± 3.2 |
31 ± 7.8 |
12 ± 3.0 |
+ |
0 (vehicle) |
132 ± 5.2 |
7 ± 3.2 |
46 ± 7.2 |
31 ± 6.1 |
16 ± 2.1 |
+ |
312.5 |
125 ± 22.4P |
7 ± 1.5P |
39 ± 2.6P |
23 ± 1.2P |
12 ± 1.5P |
+ |
625 |
123 ± 10.0P |
8 ± 2.1P |
41 ± 5.1P |
31 ± 2.5P |
14 ± 6.5P |
+ |
1250 |
125 ± 14.0P |
9 ± 3.2P |
40 ± 5.7P |
23 ± 4.5P |
14 ± 4.2P |
+ |
2500 |
116 ± 3.5P |
8 ± 2.1P |
39 ± 12.5P |
31 ± 1.2P |
17 ± 5.1P |
+ |
5000 |
121 ± 7.6P |
11 ± 3.8P |
46 ± 4.0P |
35 ± 2.5P |
13 ± 2.1P |
Positive controls, |
Name |
2AA |
2AA |
2AA |
2AA |
2AA |
Concentrations (μg/plate) |
1 |
2 |
10 |
0.5 |
2 |
|
Mean No. of colonies/plate (average of 3 ± SD) |
999 ± 117.8 |
289 ± 6.1 |
995 ± 77.9 |
420 ± 33.7 |
135 ± 13.1 |
|
AF2 = 2-(2-Furyl)-3-(5-nitro-2-furyl)acrylamide
9AA = 9-aminoacridine
2AA = 2-aminoanthracene
SA = sodium azide
P = Precipitate
Applicant's summary and conclusion
- Conclusions:
- Interpretation of results (migrated information):
negative
Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.
Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.
