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Registration Dossier
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EC number: 210-511-2 | CAS number: 617-35-6
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
EC#210-511-2 was considered to be non-mutagenic under the conditions of this test.
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- The study was conducted between 31 July 2017 and 03 September 2017.
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Qualifier:
- according to guideline
- Guideline:
- other: • Japanese Ministry of Economy, Trade and Industry, Japanese Ministry of Health, Labour and Welfare and Japanese Ministry of Agriculture, Forestry and Fisheries, 24 November 2000
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- Physical state/Appearance: Very pale yellow liquid
Batch: 61112
Purity: 98.54%
Expiry Date: 16 May 2018
Storage Conditions: Room temperature in the dark
Formulated concentrations were adjusted to allow for the stated water/impurity content (1.46%) of the test item. - Target gene:
- histidine and/or tryptophan
- 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:
- S9-mix
- Test concentrations with justification for top dose:
- Experiment 1 - Plate Incorporation Method
The maximum concentration was 5000 µg/plate (the maximum recommended dose level). Eight concentrations of the test item (1.5, 5, 15, 50, 150, 500, 1500 and 5000 g/plate) were assayed in triplicate against each tester strain, using the direct plate incorporation method.
Experiment 2 – Pre-Incubation Method
The dose range used for Experiment 2 was determined by the results of Experiment 1 and was 15, 50, 150, 500, 1500, 5000 µg/plate.
Six test item concentrations per bacterial strain were selected in Experiment 2 in order to achieve both four non toxic dose levels and the potential toxic limit of the test item following the change in test methodology from plate incorporation to pre-incubation. - Vehicle / solvent:
- The test item was immiscible in sterile distilled water at 50 mg/mL (two distinct layers observed) but was fully miscible in dimethyl sulphoxide at the same concentration in solubility checks performed in house. Dimethyl sulphoxide was therefore selected as the vehicle.
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Dimethyl sulphoxide
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- N-ethyl-N-nitro-N-nitrosoguanidine
- other:
- Remarks:
- Without S9-mix
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Dimethyl sulphoxide
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- benzo(a)pyrene
- other: 2-Aminoanthracene
- Remarks:
- with S9-mix
- Details on test system and experimental conditions:
- Test Item Preparation and Analysis
The test item was immiscible in sterile distilled water at 50 mg/mL (two distinct layers observed) but was fully miscible in dimethyl sulphoxide at the same concentration in solubility checks performed in house. Dimethyl sulphoxide was therefore selected as the vehicle.
The test item was accurately weighed and, on the day of each experiment, approximate half-log dilutions prepared in dimethyl sulphoxide by mixing on a vortex mixer. Formulated concentrations were adjusted to allow for the stated water/impurity content (1.46%) of the test item. Prior to use, the solvent was dried to remove water using molecular sieves i.e. 2 mm sodium alumino silicate pellets with a nominal pore diameter of 4 x 10-4 microns.
All formulations were used within four hours of preparation and were assumed to be stable for this period. Analysis for concentration, homogeneity and stability of the test item formulations is not a requirement of the test guidelines and was, therefore, not determined. This is an exception with regard to GLP and has been reflected in the GLP compliance statement.
Test for Mutagenicity: Experiment 1 - Plate Incorporation Method
Dose selection
The test item was tested using the following method. The maximum concentration was 5000 g/plate (the maximum recommended dose level). Eight concentrations of the test item (1.5, 5, 15, 50, 150, 500, 1500 and 5000 g/plate) were assayed in triplicate against each tester strain, using the direct plate incorporation method.
Without Metabolic Activation
0.1 mL of the appropriate concentration of test item, solvent vehicle or appropriate positive control was added to 2 mL of molten, trace amino-acid supplemented media containing 0.1 mL of one of the bacterial strain cultures and 0.5 mL of phosphate buffer. These were then mixed and overlayed onto a Vogel Bonner agar plate. Negative (untreated) controls were also performed on the same day as the mutation test. Each concentration of the test item, appropriate positive, vehicle and negative controls, and each bacterial strain, was assayed using triplicate plates.
With Metabolic Activation
The procedure was the same as described previously (see 3.3.2.2) except that following the addition of the test item formulation and bacterial culture, 0.5 mL of S9 mix was added to the molten, trace amino-acid supplemented media instead of phosphate buffer.
Incubation and Scoring
All of the plates were incubated at 37 ± 3 C for approximately 48 hours and scored for the presence of revertant colonies using an automated colony counting system. The plates were viewed microscopically for evidence of thinning (toxicity).
Test for Mutagenicity: Experiment 2 – Pre-Incubation Method
As the result of Experiment 1 was deemed negative, Experiment 2 was performed using the pre-incubation method in the presence and absence of metabolic activation.
Dose selection
The dose range used for Experiment 2 was determined by the results of Experiment 1 and was 15, 50, 150, 500, 1500, 5000 µg/plate.
Six test item concentrations per bacterial strain were selected in Experiment 2 in order to achieve both four non toxic dose levels and the potential toxic limit of the test item following the change in test methodology from plate incorporation to pre-incubation.
Without Metabolic Activation
0.1 mL of the appropriate bacterial strain culture, 0.5 mL of phosphate buffer and 0.1 mL of the test item formulation, solvent vehicle or 0.1 mL of appropriate positive control were incubated at 37 ± 3 C for 20 minutes (with shaking) prior to addition of 2 mL of molten, trace amino-acid supplemented media and subsequent plating onto Vogel Bonner plates. Negative (untreated) controls were also performed on the same day as the mutation test employing the plate incorporation method. All testing for this experiment was performed in triplicate.
With Metabolic Activation
The procedure was the same as described previously (see 3.3.3.2) except that following the addition of the test item formulation and bacterial strain culture, 0.5 mL of S9 mix was added to the tube instead of phosphate buffer, prior to incubation at 37 ± 3 C for 20 minutes (with shaking) and addition of molten, trace amino-acid supplemented media. All testing for this experiment was performed in triplicate.
Incubation and Scoring
All of the plates were incubated at 37 ± 3 C for approximately 48 hours and scored for the presence of revertant colonies using an automated colony counting system. The plates were viewed microscopically for evidence of thinning (toxicity).
Acceptability Criteria
The reverse mutation assay may be considered valid if the following criteria are met:
All bacterial strains must have demonstrated the required characteristics as determined by their respective strain checks according to Ames et al., (1975), Maron and Ames (1983) and Mortelmans and Zeiger (2000).
All tester strain cultures should exhibit a characteristic number of spontaneous revertants per plate in the vehicle and untreated controls (negative controls). Acceptable ranges are presented as follows:
TA1535 7 to 40
TA100 60 to 200
TA1537 2 to 30
TA98 8 to 60
WP2uvrA 10 to 60
All tester strain cultures should be in the range of 0.9 to 9 x 109 bacteria per mL.
Diagnostic mutagens (positive control chemicals) must be included to demonstrate both the intrinsic sensitivity of the tester strains to mutagen exposure and the integrity of the S9-mix. All of the positive control chemicals used in the study should induce marked increases in the frequency of revertant colonies, both with or without metabolic activation.
There should be a minimum of four non-toxic test item dose levels.
There should be no evidence of excessive contamination. - Evaluation criteria:
- There are several criteria for determining a positive result. Any, one, or all of the following can be used to determine the overall result of the study:
1. A dose-related increase in mutant frequency over the dose range tested (De Serres and Shelby, 1979).
2. A reproducible increase at one or more concentrations.
3. Biological relevance against in-house historical control ranges.
4. Statistical analysis of data as determined by UKEMS (Mahon et al., 1989).
5. Fold increase greater than two times the concurrent solvent control for any tester strain (especially if accompanied by an out of historical range response (Cariello and Piegorsch, 1996)).
A test item will be considered non-mutagenic (negative) in the test system if the above criteria are not met.
Although most experiments will give clear positive or negative results, in some instances the data generated will prohibit making a definite judgment about test item activity. Results of this type will be reported as equivocal. - Statistics:
- Statistical significance was confirmed by using Dunnetts Regression Analysis (* = p < 0.05) for those values that indicate statistically significant increases in the frequency of revertant colonies compared to the concurrent solvent control.
- Key result
- Species / strain:
- S. typhimurium TA 1535
- 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
- Key result
- Species / strain:
- S. typhimurium TA 1537
- 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
- Key result
- Species / strain:
- S. typhimurium TA 98
- 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
- Key result
- Species / strain:
- S. typhimurium 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
- Key result
- Species / strain:
- S. typhimurium TA 102
- 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
- Key result
- Species / strain:
- E. coli WP2 uvr A
- 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:
- Prior to use, the master strains were checked for characteristics, viability and spontaneous reversion rate (all were found to be satisfactory). The amino acid supplemented top agar and the S9-mix used in both experiments was shown to be sterile. The test item formulation was also shown to be sterile. These data are not given in the report.
Results for the negative controls (spontaneous mutation rates) were considered to be acceptable. These data are for concurrent untreated control plates performed on the same day as the Mutation Test.
The vehicle (dimethyl sulphoxide) control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9-mix were validated.
The maximum dose level of the test item in the first experiment was selected as the maximum recommended dose level of 5000 µg/plate. There was no visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation (S9-mix), in the first mutation test (plate incorporation method). Consequently, the same maximum dose level was used as the maximum concentration in the second mutation test. Similarly, there was no visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation (S9-mix), in the second mutation test (pre-incubation method).
No test item precipitate was observed on the plates at any of the doses tested in either the presence or absence of S9-mix.
There were no significant increases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 1 (plate incorporation method). Similarly, no significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 2 (pre incubation method). - Conclusions:
- EC#210-511-2 was considered to be non-mutagenic under the conditions of this test.
- Executive summary:
Introduction
The test method was designed to be compatible with the guidelines for bacterial mutagenicity testing published by the major Japanese Regulatory Authorities including METI, MHLW and MAFF, the OECD Guidelines for Testing of Chemicals No. 471 "Bacterial Reverse Mutation Test", Method B13/14 of Commission Regulation (EC) number 440/2008 of 30 May 2008 and the USA, EPA OCSPP harmonized guideline - Bacterial Reverse Mutation Test.
Methods
Salmonella typhimuriumstrains TA1535, TA1537, TA98 and TA100 andEscherichia colistrain WP2uvrAwere treated with the test item using both the Ames plate incorporation and pre-incubation methods at up to eight dose levels, in triplicate, both with and without the addition of a rat liver homogenate metabolizing system (10% liver S9 in standard co-factors). The dose range for Experiment 1 was predetermined and was 1.5 to 5000 mg/plate. The experiment was repeated on a separate day (pre-incubation method) using fresh cultures of the bacterial strains and fresh test item formulations. The dose range was amended following the results of Experiment 1 and was 15 to 5000 µg/plate. Six test item concentrations were selected in Experiment 2 in order to achieve both four non‑toxic dose levels and the potential toxic limit of the test item following the change in test methodology.
Results
The vehicle (dimethyl sulphoxide) control plates gave counts of revertant colonies within the normal range. All of the positive control chemicals used in the test induced marked increases in the frequency of revertant colonies, both with or without metabolic activation. Thus, the sensitivity of the assay and the efficacy of the S9-mix were validated.
The maximum dose level of the test item in the first experiment was selected as the maximum recommended dose level of 5000 µg/plate. There was no visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation (S9-mix), in the first mutation test (plate incorporation method). Consequently, the same maximum dose level was used as the maximum concentration in the second mutation test. Similarly, there was no visible reduction in the growth of the bacterial background lawn at any dose level, either in the presence or absence of metabolic activation (S9-mix), in the second mutation test (pre-incubation method).
No test item precipitate was observed on the plates at any of the doses tested in either the presence or absence of S9-mix.
There were no significantincreases in the frequency of revertant colonies recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 1 (plate incorporation method). Similarly, no significant increases in the frequency of revertant colonies were recorded for any of the bacterial strains, with any dose of the test item, either with or without metabolic activation (S9-mix) in Experiment 2 (pre‑incubation method).
Conclusion
EC#210-511-2 was considered to be non-mutagenic under the conditions of this test.
Reference
Spontaneous Mutation Rates (Concurrent Negative Controls)
Experiment 1
Number of revertants (mean number of colonies per plate) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
|||||
81 |
|
16 |
|
34 |
|
27 |
|
19 |
|
79 |
(84) |
16 |
(17) |
30 |
(32) |
20 |
(24) |
15 |
(16) |
92 |
|
20 |
|
32 |
|
25 |
|
15 |
|
Experiment 2
Number of revertants (mean number of colonies per plate) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
|||||
79 |
|
25 |
|
22 |
|
27 |
|
9 |
|
74 |
(72) |
20 |
(23) |
20 |
(21) |
15 |
(20) |
6 |
(11) |
63 |
|
24 |
|
21 |
|
19 |
|
17 |
|
Test Results: Experiment 1 – Without Metabolic Activation
Test Period |
From: 15 August 2017 |
To: 18 August 2017 |
||||||||||
S9-Mix (-) |
Dose Level Per Plate |
Number of revertants (mean) +/- SD |
||||||||||
Base-pair substitution strains |
Frameshift strains |
|||||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||||||||
Solvent Control (DMSO) |
73 68 77 |
(73) 4.5# |
26 19 21 |
(22) 3.6 |
16 25 16 |
(19) 5.2 |
23 19 20 |
(21) 2.1 |
4 12 5 |
(7) 4.4 |
||
1.5 µg |
71 76 83 |
(77) 6.0 |
26 25 17 |
(23) 4.9 |
26 25 15 |
(22) 6.1 |
25 29 28 |
(27) 2.1 |
13 9 7 |
(10) 3.1 |
||
5 µg |
75 74 88 |
(79) 7.8 |
29 25 22 |
(25) 3.5 |
20 25 34 |
(26) 7.1 |
26 23 17 |
(22) 4.6 |
6 8 7 |
(7) 1.0 |
||
15 µg |
75 92 82 |
(83) 8.5 |
26 17 16 |
(20) 5.5 |
22 17 24 |
(21) 3.6 |
20 14 20 |
(18) 3.5 |
11 8 8 |
(9) 1.7 |
||
50 µg |
70 75 83 |
(76) 6.6 |
20 27 22 |
(23) 3.6 |
23 15 14 |
(17) 4.9 |
27 17 13 |
(19) 7.2 |
15 7 9 |
(10) 4.2 |
||
150 µg |
117 73 73 |
(88) 25.4 |
21 23 26 |
(23) 2.5 |
32 18 19 |
(23) 7.8 |
17 30 21 |
(23) 6.7 |
19 10 8 |
(12) 5.9 |
||
500 µg |
71 72 78 |
(74) 3.8 |
19 15 19 |
(18) 2.3 |
13 12 18 |
(14) 3.2 |
27 23 15 |
(22) 6.1 |
11 13 15 |
(13) 2.0 |
||
1500 µg |
64 75 86 |
(75) 11.0 |
20 25 19 |
(21) 3.2 |
14 20 29 |
(21) 7.5 |
25 23 24 |
(24) 1.0 |
6 6 4 |
(5) 1.2 |
||
5000 µg |
77 88 97 |
(87) 10.0 |
22 27 15 |
(21) 6.0 |
29 22 17 |
(23) 6.0 |
29 20 11 |
(20) 9.0 |
14 13 10 |
(12) 2.1 |
||
Positive controls S9-Mix (-) |
Name Dose Level No. of Revertants |
ENNG |
ENNG |
ENNG |
4NQO |
9AA |
||||||
3 µg |
5 µg |
2 µg |
0.2 µg |
80 µg |
||||||||
543 574 490 |
(536) 42.5 |
371 519 462 |
(451) 74.6 |
301 272 245 |
(273) 28.0 |
213 195 202 |
(203) 9.1 |
347 330 189 |
(289) 86.7 |
|||
Test Results: Experiment 1 – With Metabolic Activation
Test Period |
From: 15 August 2017 |
To: 18 August 2017 |
||||||||||
S9-Mix (+) |
Dose Level Per Plate |
Number of revertants (mean) +/- SD |
||||||||||
Base-pair substitution strains |
Frameshift strains |
|||||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||||||||
Solvent Control (DMSO) |
79 69 78 |
(75) 5.5# |
22 20 19 |
(20) 1.5 |
29 21 24 |
(25) 4.0 |
30 28 29 |
(29) 1.0 |
25 10 7 |
(14) 9.6 |
||
1.5 µg |
76 76 85 |
(79) 5.2 |
12 9 22 |
(14) 6.8 |
35 31 30 |
(32) 2.6 |
24 37 31 |
(31) 6.5 |
7 24 9 |
(13) 9.3 |
||
5 µg |
89 66 80 |
(78) 11.6 |
16 17 15 |
(16) 1.0 |
30 21 22 |
(24) 4.9 |
22 28 23 |
(24) 3.2 |
14 10 10 |
(11) 2.3 |
||
15 µg |
92 86 86 |
(88) 3.5 |
20 17 22 |
(20) 2.5 |
31 30 31 |
(31) 0.6 |
27 23 32 |
(27) 4.5 |
11 8 11 |
(10) 1.7 |
||
50 µg |
73 74 82 |
(76) 4.9 |
19 13 16 |
(16) 3.0 |
28 25 29 |
(27) 2.1 |
15 25 23 |
(21) 5.3 |
6 9 16 |
(10) 5.1 |
||
150 µg |
91 76 84 |
(84) 7.5 |
14 23 23 |
(20) 5.2 |
22 33 26 |
(27) 5.6 |
26 27 35 |
(29) 4.9 |
10 17 16 |
(14) 3.8 |
||
500 µg |
74 69 74 |
(72) 2.9 |
22 21 20 |
(21) 1.0 |
22 28 26 |
(25) 3.1 |
26 29 15 |
(23) 7.4 |
17 12 20 |
(16) 4.0 |
||
1500 µg |
88 72 66 |
(75) 11.4 |
17 22 16 |
(18) 3.2 |
28 30 19 |
(26) 5.9 |
36 28 20 |
(28) 8.0 |
11 13 9 |
(11) 2.0 |
||
5000 µg |
72 86 80 |
(79) 7.0 |
16 27 28 |
(24) 6.7 |
17 17 27 |
(20) 5.8 |
24 24 23 |
(24) 0.6 |
16 6 11 |
(11) 5.0 |
||
Positive controls S9-Mix (+) |
Name Dose Level No. of Revertants |
2AA |
2AA |
2AA |
BP |
2AA |
||||||
1 µg |
2 µg |
10 µg |
5 µg |
2 µg |
||||||||
1296 1604 1451 |
(1450) 154.0 |
275 288 299 |
(287) 12.0 |
165 148 106 |
(140) 30.4 |
359 352 192 |
(301) 94.5 |
498 592 520 |
(537) 49.2 |
|||
Test Results: Experiment 2 – Without Metabolic Activation
Test Period |
From: 31 August 2017 |
To: 03 September 2017 |
||||||||||
S9-Mix (-) |
Dose Level Per Plate |
Number of revertants (mean) +/- SD |
||||||||||
Base-pair substitution strains |
Frameshift strains |
|||||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||||||||
Solvent Control (DMSO) |
65 83 74 |
(74) 9.0# |
27 26 28 |
(27) 1.0 |
16 19 19 |
(18) 1.7 |
24 19 20 |
(21) 2.6 |
14 10 11 |
(12) 2.1 |
||
15 µg |
79 64 85 |
(76) 10.8 |
27 26 21 |
(25) 3.2 |
27 21 31 |
(26) 5.0 |
24 16 21 |
(20) 4.0 |
19 16 12 |
(16) 3.5 |
||
50 µg |
80 74 65 |
(73) 7.5 |
24 25 26 |
(25) 1.0 |
10 18 18 |
(15) 4.6 |
20 29 29 |
(26) 5.2 |
10 12 16 |
(13) 3.1 |
||
150 µg |
66 84 85 |
(78) 10.7 |
27 28 24 |
(26) 2.1 |
23 12 20 |
(18) 5.7 |
20 30 19 |
(23) 6.1 |
19 12 16 |
(16) 3.5 |
||
500 µg |
83 80 73 |
(79) 5.1 |
26 26 32 |
(28) 3.5 |
24 10 25 |
(20) 8.4 |
18 18 20 |
(19) 1.2 |
6 11 7 |
(8) 2.6 |
||
1500 µg |
88 57 81 |
(75) 16.3 |
26 25 28 |
(26) 1.5 |
29 27 28 |
(28) 1.0 |
19 23 15 |
(19) 4.0 |
19 9 15 |
(14) 5.0 |
||
5000 µg |
89 77 94 |
(87) 8.7 |
30 29 29 |
(29) 0.6 |
17 24 22 |
(21) 3.6 |
28 16 18 |
(21) 6.4 |
13 10 9 |
(11) 2.1 |
||
Positive controls S9-Mix (-) |
Name Dose Level No. of Revertants |
ENNG |
ENNG |
ENNG |
4NQO |
9AA |
||||||
3 µg |
5 µg |
2 µg |
0.2 µg |
80 µg |
||||||||
1050 1180 1009 |
(1080) 89.3 |
2329 2189 1900 |
(2139) 218.8 |
1167 1141 1089 |
(1132) 39.7 |
340 333 186 |
(286) 87.0 |
374 405 260 |
(346) 76.4 |
|||
ENNG4NQO9AA#
Table5 Test Results: Experiment 2 – With Metabolic Activation
Test Period |
From: 31 August 2017 |
To: 03 September 2017 |
||||||||||
S9-Mix (+) |
Dose Level Per Plate |
Number of revertants (mean) +/- SD |
||||||||||
Base-pair substitution strains |
Frameshift strains |
|||||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||||||||
Solvent Control (DMSO) |
68 73 62 |
(68) 5.5# |
38 28 32 |
(33) 5.0 |
28 17 27 |
(24) 6.1 |
27 22 20 |
(23) 3.6 |
14 17 16 |
(16) 1.5 |
||
15 µg |
65 73 64 |
(67) 4.9 |
26 33 30 |
(30) 3.5 |
32 21 23 |
(25) 5.9 |
25 24 35 |
(28) 6.1 |
15 15 8 |
(13) 4.0 |
||
50 µg |
56 63 73 |
(64) 8.5 |
29 33 28 |
(30) 2.6 |
29 28 24 |
(27) 2.6 |
22 14 28 |
(21) 7.0 |
21 9 10 |
(13) 6.7 |
||
150 µg |
60 61 56 |
(59) 2.6 |
28 30 32 |
(30) 2.0 |
29 17 21 |
(22) 6.1 |
16 26 26 |
(23) 5.8 |
12 13 14 |
(13) 1.0 |
||
500 µg |
65 63 64 |
(64) 1.0 |
30 26 29 |
(28) 2.1 |
20 17 21 |
(19) 2.1 |
21 21 31 |
(24) 5.8 |
6 16 9 |
(10) 5.1 |
||
1500 µg |
68 72 58 |
(66) 7.2 |
30 27 27 |
(28) 1.7 |
24 32 26 |
(27) 4.2 |
23 25 22 |
(23) 1.5 |
11 16 13 |
(13) 2.5 |
||
5000 µg |
80 83 73 |
(79) 5.1 |
28 25 33 |
(29) 4.0 |
23 26 30 |
(26) 3.5 |
25 30 23 |
(26) 3.6 |
11 11 9 |
(10) 1.2 |
||
Positive controls S9-Mix (+) |
Name Dose Level No. of Revertants |
2AA |
2AA |
2AA |
BP |
2AA |
||||||
1 µg |
2 µg |
10 µg |
5 µg |
2 µg |
||||||||
1320 1270 1324 |
(1305) 30.1 |
427 371 422 |
(407) 31.0 |
166 164 177 |
(169) 7.0 |
164 178 154 |
(165) 12.1 |
511 459 460 |
(477) 29.7 |
|||
ENNG N-ethyl-N'-nitro-N-nitrosoguanidine
4NQO 4-Nitroquinoline-1-oxide
9AA 9-Aminoacridine
# Standard deviation
BP Benzo(a)pyrene
2AA 2-AminoanthraceneEndpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Justification for classification or non-classification
The test substance was considered to be non-mutagenic under the Ames study therefore the substance is not classified for Genetic toxicity.
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.
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