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EC number: 946-923-0 | CAS number: -
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Endpoint summary
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Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
The substance was found to be non mutagenic in Ames test (OECD 471, GLP, K1).
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:
- May - October 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)
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- histidine locus for Salmonella strains and tryptophan for E. coli strain
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Details on mammalian cell type (if applicable):
- not applicable
- Additional strain / cell type characteristics:
- not applicable
- Cytokinesis block (if used):
- not applicable
- Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver homogenate metabolizing system (10% liver S9 in standard co-factors)
- Test concentrations with justification for top dose:
- Experiment 1 - Plate Incorporation Method: 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate, with and without S9-mix
Experiment 2 - Pre-Incubation Method: 0.5, 1.5, 5, 15, 50, 150, 500 µg/plate, without S9-mix for TA100, TA1535 and TA 1537 and 5, 15, 50, 150, 500, 1500 and 5000 µg/plate for TA98 without S9. 15, 50, 150, 500, 1500and 5000 μg/plate with S9 for all strains. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: Acetone
In solubility checks performed in house the test item was noted as immiscible in sterile distilled water and dimethyl sulphoxide at 50 mg/mL but fully miscible in acetone at 100 mg/mL. Acetone was therefore selected as the vehicle. - Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Acetone
- Positive controls:
- yes
- Positive control substance:
- 4-nitroquinoline-N-oxide
- 9-aminoacridine
- N-ethyl-N-nitro-N-nitrosoguanidine
- benzo(a)pyrene
- other: 2-Aminoanthracene (2AA)
- Details on test system and experimental conditions:
- SOURCE OF TEST SYSTEM: The bacteria used in the test were obtained from:
- University of California, Berkeley, on culture discs, on 04 August 1995
- British Industrial Biological Research Association, on a nutrient agar plate, on 17 August 1987
METHOD OF APPLICATION: preincubation
All testing will be performed using the pre-incubation method (20 minutes at 37 °C) except for the untreated controls.
DURATION
- Preincubation period: 20 minutes.
- Exposure duration: ca. 48 hours
CONTROLS:
- Vehicle/solvent control: Acetone
- Negative (untreated) controls were performed to assess the spontaneous revertant colony rate.
- Positive control items used demonstrated a direct and indirect acting mutagenic effect depending on the presence or absence of metabolic activation.
- Sterility controls were performed in triplicate as follows:
Top agar and histidine/biotin or tryptophan in the absence of S9-mix;
Top agar and histidine/biotin or tryptophan in the presence of S9-mix; and
The maximum dosing solution of the test item in the absence of S9-mix only (test in singular only).
NUMBER OF REPLICATIONS: Triplicate
- OTHER: 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). Manual counts were performed at 5000 µg/plate because of test item precipitation. Several further manual counts were also required due to revertant colonies spreading slightly, thus distorting the actual plate count. - Rationale for test conditions:
- The dose range for Experiment 1 was predetermined and was 1.5 to 5000 µg/plate (i.e. maximum recommended dose level). The dose range used for Experiment 2 was determined by the results of Experiment 1. Up to seven test item dose levels per bacterial strain were selected in the second mutation test in order to achieve both a minimum of four non-toxic dose levels and the toxic limit of the test item.
- Evaluation criteria:
- Criteria for determining a positive result:
- A dose-related increase in mutant frequency over the dose range tested (De Serres and Shelby, 1979).
- A reproducible increase at one or more concentrations.
- Biological relevance against in-house historical control ranges.
- 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. - Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- 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:
- 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:
- 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:
- cytotoxicity
- Vehicle 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:
- cytotoxicity
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: A test item precipitate (light and globular in appearance) was noted at 5000 µg/plate, this observation did not prevent the scoring of revertant colonies.
MUTAGENICITY
- The vehicle (acetone) 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.
- In the first mutation test, the test item caused a visible reduction in the growth of the bacterial background lawns of all of the bacterial tester strains from 150 µg/plate (TA100, TA 1535 and TA 1537) and 1500 µg/plate (TA98 and WP2uvrA) in the absence of S9 mix and to two bacterial strains (TA100 and TA1535) at 5000 µg/plate in the presence S9-mix. In the second mutation test, the test item again induced a toxic response with weakened bacterial background lawns noted in the absence of S9-mix from 150 µg/plate (TA1535, TA100 and TA1537) and 1500 µg/plate (TA98 and WP2uvrA). In the presence of S9-mix, weakened bacterial background lawns were noted to three of the bacterial tester strains at 5000 µg/plate (TA100, TA1535 and WP2uvrA).
- 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 1 and Experiment 2 .
- Refer Tables 7.6.1/1 to 7.6.1/5 for more details.
HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: Refer Table 7.6.1/6
- Negative (solvent/vehicle) historical control data: Refer Table 7.6.1/6
OTHERS:
- 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.
- Results for the negative controls (spontaneous mutation rates) are presented in 7.6.1/1 and were considered to be acceptable. These data are for concurrent untreated control plates performed on the same day as the Mutation Test. - Conclusions:
- Under the test conditions, the test item is not considered as mutagenic in Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100, and Escherichia coli strain WP2uvrA.
- Executive summary:
In a reverse gene mutation assay in bacteria, performed according to the OECD Guideline 471 and in compliance with GLP, Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100, and Escherichia coli strain WP2uvrA were exposed to the test item at the following concentrations:
- Experiment 1 - Pre incubation Method: 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate, with and without S9-mix
- Experiment 2 - Pre-Incubation Method: 0.5, 1.5, 5, 15, 50, 150, 500 µg/plate, without S9-mix for TA100, TA1535 and TA 1537 and 5, 15, 50, 150, 500, 1500 and 5000 µg/plate for TA98 without S9. 15, 50, 150, 500, 1500and 5000 μg/plate with S9 for all strains.
Rat liver homogenate (10% liver S9 in standard co-factors) was used as a metabolizing system. Vehicle control, negative (untreated) and positive control groups were also included in mutagenicity tests.
The vehicle (acetone) 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.
In the first mutation test, the test item caused a visible reduction in the growth of the bacterial background lawns of all of the bacterial tester strains from 150 µg/plate (TA100, TA 1535 and TA 1537) and 1500 µg/plate (TA98 and WP2uvrA) in the absence of S9 mix and to two bacterial strains (TA100 and TA1535) at 5000 µg/plate in the presence S9-mix. In the second mutation test, the test item again induced a toxic response with weakened bacterial background lawns noted in the absence of S9-mix from 150 µg/plate (TA1535, TA100 and TA1537) and 1500 µg/plate (TA98 and WP2uvrA). In the presence of S9-mix, weakened bacterial background lawns were noted to three of the bacterial tester strains at 5000 µg/plate (TA100, TA1535 and WP2uvrA).
A test item precipitate (light and globular in appearance) was noted at 5000 µg/plate, this observation did not prevent the scoring of revertant colonies.
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 1 and Experiment 2 .
Under the test conditions, the test item is not considered as mutagenic in these bacterial systems.
Reference
Table 7.6.1/1:Spontaneous Mutation Rates (Concurrent Negative Controls)
Number of revertants (mean number of colonies per plate) |
|||||||||
Base-pair substitution type |
Frameshift type |
||||||||
Experiment 1 |
|||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
|||||
87 |
|
16 |
|
31 |
|
31 |
|
9 |
|
86 |
(84) |
19 |
(17) |
15 |
(22) |
15 |
(24) |
21 |
(15) |
80 |
|
17 |
|
19 |
|
25 |
|
16 |
|
Experiment 2 |
|||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
|||||
65 |
|
18 |
|
36 |
|
20 |
|
15 |
|
65 |
(64) |
23 |
(18) |
32 |
(34) |
15 |
(18) |
18 |
(16) |
61 |
|
14 |
|
34 |
|
19 |
|
16 |
|
Table 7.6.1/2:Test Results: Experiment 1 – Without Metabolic Activation
Test Period |
From: 25 July 2017 |
To: 28 July 2018 |
||||||||||
S9-Mix (-) |
Dose Level Per Plate |
Number of revertants (mean) +/- SD |
||||||||||
Base-pair substitution strains |
Frameshift strains |
|||||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||||||||
Solvent Control (Acetone) |
83 81 88 |
(84) 3.6# |
26 14 24 |
(21) 6.4 |
20 28 25 |
(24) 4.0 |
26 16 11 |
(18) 7.6 |
15 21 19 |
(18) 3.1 |
||
1.5 µg |
90 71 84 |
(82) 9.7 |
9 16 12 |
(12) 3.5 |
25 19 24 |
(23) 3.2 |
15 21 25 |
(20) 5.0 |
18 8 16 |
(14) 5.3 |
||
5 µg |
76 71 85 |
(77) 7.1 |
8 21 15 |
(15) 6.5 |
16 25 20 |
(20) 4.5 |
25 16 14 |
(18) 5.9 |
8 13 8 |
(10) 2.9 |
||
15 µg |
96 94 111 |
(100) 9.3 |
12 16 9 |
(12) 3.5 |
18 24 32 |
(25) 7.0 |
10 21 17 |
(16) 5.6 |
9 9 14 |
(11) 2.9 |
||
50 µg |
84 82 93 |
(86) 5.9 |
9 9 8 |
(9) 0.6 |
28 25 27 |
(27) 1.5 |
16 16 21 |
(18) 2.9 |
15 9 21 |
(15) 6.0 |
||
150 µg |
86 S 79 S 80 S |
(82) 3.8 |
8 S 6 S 20 S |
(11) 7.6 |
25 28 12 |
(22) 8.5 |
15 13 18 |
(15) 2.5 |
3 S 3 S 3 S |
(3) 0.0 |
||
500 µg |
74 S 65 S 58 S |
(66) 8.0 |
10 S 12 S 8 S |
(10) 2.0 |
24 26 20 |
(23) 3.1 |
27 23 17 |
(22) 5.0 |
7 S 6 S 3 S |
(5) 2.1 |
||
1500 µg |
16 S 15 S 26 S |
(19) 6.1 |
14 S 5 S 12 S |
(10) 4.7 |
23 S 20 S 8 S |
(17) 7.9 |
12 S 12 S 14 S |
(13) 1.2 |
3 S 2 S 2 S |
(2) 0.6 |
||
5000 µg |
0 VP 0 VP 0 VP |
(0) 0.0 |
0 VP 0 VP 0 VP |
(0) 0.0 |
14 SP 14 SP 13 SP |
(14) 0.6 |
18 SP 18 SP 17 SP |
(18) 0.6 |
0 VP 0 VP 0 VP |
(0) 0.0 |
||
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 |
||||||||
1692 1753 1736 |
(1727) 31.5 |
596 637 489 |
(574) 76.4 |
719 710 811 |
(747) 55.9 |
311 277 286 |
(291) 17.6 |
251 111 175 |
(179) 70.1 |
ENNG:N-ethyl-N'-nitro-N-nitrosoguanidine
4NQO:4-Nitroquinoline-1-oxide
9AA: 9-Aminoacridine
P: Test item precipitate
#: Standard deviation
Table 7.6.1/3:Test Results: Experiment 1 – With Metabolic Activation
Test Period |
From: 25 July 2017 |
To: 28 July 2018 |
||||||||||
S9-Mix (+) |
Dose Level Per Plate |
Number of revertants (mean) +/- SD |
||||||||||
Base-pair substitution strains |
Frameshift strains |
|||||||||||
TA100 |
TA1535 |
WP2uvrA |
TA98 |
TA1537 |
||||||||
Solvent Control (Acetone) |
89 82 90 |
(87) 4.4# |
15 17 16 |
(16) 1.0 |
32 20 41 |
(31) 10.5 |
29 40 32 |
(34) 5.7 |
15 17 20 |
(17) 2.5 |
||
1.5 µg |
74 85 78 |
(79) 5.6 |
10 10 18 |
(13) 4.6 |
21 20 31 |
(24) 6.1 |
34 16 29 |
(26) 9.3 |
12 11 17 |
(13) 3.2 |
||
5 µg |
97 76 91 |
(88) 10.8 |
18 15 14 |
(16) 2.1 |
24 23 30 |
(26) 3.8 |
31 44 32 |
(36) 7.2 |
12 10 12 |
(11) 1.2 |
||
15 µg |
81 82 78 |
(80) 2.1 |
14 18 24 |
(19) 5.0 |
18 21 25 |
(21) 3.5 |
13 25 30 |
(23) 8.7 |
21 21 20 |
(21) 0.6 |
||
50 µg |
68 90 94 |
(84) 14.0 |
20 13 12 |
(15) 4.4 |
31 31 33 |
(32) 1.2 |
25 23 31 |
(26) 4.2 |
14 12 20 |
(15) 4.2 |
||
150 µg |
96 95 84 |
(92) 6.7 |
12 21 14 |
(16) 4.7 |
34 26 24 |
(28) 5.3 |
35 36 27 |
(33) 4.9 |
17 13 17 |
(16) 2.3 |
||
500 µg |
92 72 105 |
(90) 16.6 |
9 23 15 |
(16) 7.0 |
31 33 19 |
(28) 7.6 |
25 28 23 |
(25) 2.5 |
21 20 13 |
(18) 4.4 |
||
1500 µg |
90 82 84 |
(85) 4.2 |
16 26 12 |
(18) 7.2 |
29 25 24 |
(26) 2.6 |
23 29 33 |
(28) 5.0 |
12 13 21 |
(15) 4.9 |
||
5000 µg |
64 SP 66 SP 71 SP |
(67) 3.6 |
10 SP 14 SP 10 SP |
(11) 2.3 |
39 P 18 P 15 P |
(24) 13.1 |
23 P 34 P 20 P |
(26) 7.4 |
7 P 11 P 11 P |
(10) 2.3 |
||
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 |
||||||||
1170 1028 1286 |
(1161) 129.2 |
288 320 285 |
(298) 19.4 |
168 168 166 |
(167) 1.2 |
144 158 170 |
(157) 13.0 |
467 454 475 |
(465) 10.6 |
2AA: 2-Aminoanthracene
BP: Benzo(a)pyrene
P: Test item precipitate
#: Standard deviation
Table 7.6.1/4:Test Results: Experiment 2 – Without Metabolic Activation
Test Period |
From: 14 August 2017 |
To: 17 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 (Acetone) |
78 65 61 |
(68) 8.9# |
19 19 20 |
(19) 0.6 |
20 23 23 |
(22) 1.7 |
20 21 22 |
(21) 1.0 |
19 16 5 |
(13) 7.4 |
||
0.5 µg |
64 70 83 |
(72) 9.7 |
17 19 14 |
(17) 2.5 |
N/T |
N/T |
19 11 14 |
(15) 4.0 |
||||
1.5 µg |
79 78 63 |
(73) 9.0 |
13 23 19 |
(18) 5.0 |
N/T |
N/T |
13 17 5 |
(12) 6.1 |
||||
5 µg |
88 65 74 |
(76) 11.6 |
22 21 16 |
(20) 3.2 |
20 20 29 |
(23) 5.2 |
13 25 21 |
(20) 6.1 |
16 9 11 |
(12) 3.6 |
||
15 µg |
62 71 73 |
(69) 5.9 |
22 17 16 |
(18) 3.2 |
28 33 28 |
(30) 2.9 |
20 28 29 |
(26) 4.9 |
13 18 15 |
(15) 2.5 |
||
50 µg |
64 68 74 |
(69) 5.0 |
18 31 18 |
(22) 7.5 |
29 18 21 |
(23) 5.7 |
17 20 25 |
(21) 4.0 |
9 9 10 |
(9) 0.6 |
||
150 µg |
63 S 67 S 60 S |
(63) 3.5 |
12 S 17 S 21 S |
(17) 4.5 |
28 19 22 |
(23) 4.6 |
29 15 17 |
(20) 7.6 |
10 S 5 S 14 S |
(10) 4.5 |
||
500 µg |
58 S 41 S 51 S |
(50) 8.5 |
14 S 15 S 13 S |
(14) 1.0 |
21 20 26 |
(22) 3.2 |
14 16 12 |
(14) 2.0 |
5 S 7 S 6 S |
(6) 1.0 |
||
1500 µg |
N/T |
N/T |
17 S 17 S 21 S |
(18) 2.3 |
14 S 12 S 26 S |
(17) 7.6 |
N/T |
|||||
5000 µg |
N/T |
N/T |
31 SP 20 SP 28 SP |
(26) 5.7 |
9 SP 11 SP 11 SP |
(10) 1.2 |
N/T |
|||||
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 |
||||||||
807 620 724 |
(717) 93.7 |
1500 1528 1598 |
(1542) 50.5 |
668 627 485 |
(593) 96.0 |
175 205 180 |
(187) 16.1 |
347 152 193 |
(231) 102.8 |
ENNG:N-ethyl-N'-nitro-N-nitrosoguanidine
4NQO:4-Nitroquinoline-1-oxide
9AA: 9-Aminoacridine
P: Test item precipitate
#: Standard deviation
Table 7.6.1/5:Test Results: Experiment 2 – With Metabolic Activation
Test Period |
From: 14 August 2017 |
To: 17 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 (Acetone) |
71 67 85 |
(74) 9.5# |
16 20 22 |
(19) 3.1 |
35 24 33 |
(31) 5.9 |
34 24 38 |
(32) 7.2 |
13 14 7 |
(11) 3.8 |
||
15 µg |
61 75 89 |
(75) 14.0 |
26 22 22 |
(23) 2.3 |
40 36 31 |
(36) 4.5 |
26 32 39 |
(32) 6.5 |
13 15 13 |
(14) 1.2 |
||
50 µg |
74 75 76 |
(75) 1.0 |
21 24 13 |
(19) 5.7 |
29 41 33 |
(34) 6.1 |
40 36 24 |
(33) 8.3 |
17 13 14 |
(15) 2.1 |
||
150 µg |
86 82 75 |
(81) 5.6 |
11 18 29 |
(19) 9.1 |
24 45 31 |
(33) 10.7 |
36 36 26 |
(33) 5.8 |
20 15 15 |
(17) 2.9 |
||
500 µg |
79 79 75 |
(78) 2.3 |
17 17 33 |
(22) 9.2 |
27 34 31 |
(31) 3.5 |
44 13 42 |
(33) 17.3 |
11 18 21 |
(17) 5.1 |
||
1500 µg |
62 65 65 |
(64) 1.7 |
22 21 31 |
(25) 5.5 |
33 35 26 |
(31) 4.7 |
23 32 33 |
(29) 5.5 |
11 13 8 |
(11) 2.5 |
||
5000 µg |
64 SP 60 SP 55 SP |
(60) 4.5 |
26 SP 31 SP 29 SP |
(29) 2.5 |
19 SP 23 SP 23 SP |
(22) 2.3 |
30 P 29 P 34 P |
(31) 2.6 |
15 P 19 P 18 P |
(17) 2.1 |
||
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 |
||||||||
1029 1096 1037 |
(1054) 36.6 |
233 212 200 |
(215) 16.7 |
139 117 157 |
(138) 20.0 |
137 107 70 |
(105) 33.6 |
394 453 413 |
(420) 30.1 |
2AA 2-Aminoanthracene
BP Benzo(a)pyrene
P Test item precipitate
S Sparse bacterial background lawn
# Standard deviation
Table 7.6.1/6:History Profile of Vehicle and Positive Control Values
OMBINED VEHICLE AND UNTREATED CONTROL VALUES 2015 |
|||||||||||||||||||||||||||||||||
Strain S9-Mix |
TA100 |
TA1535 |
TA102 |
WP2uvrA |
TA98 |
TA1537 |
WP2uvrA pKM101 |
WP2pKM101 |
|||||||||||||||||||||||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
||||||||||||||||||
Values† |
274 |
278 |
504 |
285 |
26 |
13 |
461 |
229 |
526 |
299 |
506 |
282 |
42 |
51 |
39 |
49 |
|||||||||||||||||
Min |
60 |
61 |
7 |
7 |
222 |
278 |
10 |
12 |
11 |
10 |
4 |
6 |
87 |
98 |
89 |
93 |
|||||||||||||||||
Max |
166 |
175 |
31 |
29 |
376 |
388 |
58 |
43 |
45 |
46 |
27 |
27 |
237 |
254 |
174 |
177 |
|||||||||||||||||
Mean |
91 |
95 |
16 |
14 |
286 |
333 |
24 |
27 |
21 |
24 |
12 |
13 |
156 |
164 |
123 |
137 |
|||||||||||||||||
SD |
19.3 |
19.1 |
4.5 |
4.0 |
48.7 |
37.6 |
5.6 |
5.9 |
6.2 |
6.1 |
3.8 |
3.4 |
42.2 |
35.6 |
23.1 |
21.2 |
|||||||||||||||||
POSITIVE CONTROL VALUES 2015 |
|
||||||||||||||||||||||||||||||||
Strain S9-Mix |
TA100 |
TA1535 |
TA102 |
WP2uvrA |
TA98 |
TA1537 |
WP2uvrA pKM101 |
WP2pKM101 |
|
||||||||||||||||||||||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
|||||||||||||||||
Values |
276 |
280 |
252 |
264 |
13 |
13 |
231 |
227 |
262 |
276 |
253 |
261 |
20 |
35 |
20 |
35 |
|
||||||||||||||||
Min |
222 |
250 |
79 |
118 |
953 |
673 |
116 |
103 |
100 |
78 |
164 |
97 |
430 |
494 |
745 |
325 |
|
||||||||||||||||
Max |
2266 |
2402 |
2779 |
457 |
3140 |
1655 |
2769 |
550 |
502 |
705 |
2318 |
823 |
1696 |
2264 |
3662 |
1174 |
|
||||||||||||||||
Mean |
614 |
927 |
472 |
246 |
2303 |
1093 |
792 |
266 |
222 |
218 |
911 |
336 |
761 |
1461 |
2257 |
569 |
|
||||||||||||||||
SD |
260.6 |
452.5 |
434.8 |
55.7 |
815.2 |
376.5 |
342.1 |
97.7 |
70.2 |
107.6 |
412.4 |
135.7 |
350.0 |
382.0 |
790.7 |
220.3 |
|
||||||||||||||||
COMBINED VEHICLE AND UNTREATED CONTROL VALUES 2016 |
|||||||||||||||||||||||||||||||||
Strain S9-Mix |
TA100 |
TA1535 |
TA102 |
WP2uvrA |
TA98 |
TA1537 |
WP2uvrA pKM101 |
WP2pKM101 |
|||||||||||||||||||||||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
||||||||||||||||||
Values |
399 |
401 |
758 |
393 |
60 |
30 |
690 |
345 |
788 |
415 |
762 |
398 |
32 |
32 |
16 |
24 |
|||||||||||||||||
Min |
63 |
66 |
8 |
8 |
216 |
221 |
10 |
13 |
8 |
12 |
3 |
4 |
97 |
104 |
78 |
52 |
|||||||||||||||||
Max |
154 |
156 |
34 |
39 |
340 |
375 |
53 |
53 |
49 |
51 |
24 |
23 |
268 |
243 |
148 |
166 |
|||||||||||||||||
Mean |
90 |
93 |
15 |
15 |
268 |
310 |
22 |
27 |
21 |
25 |
12 |
13 |
161 |
159 |
118 |
110 |
|||||||||||||||||
SD |
14.5 |
14.3 |
4.5 |
5.2 |
26.4 |
31.1 |
5.8 |
6.3 |
4.8 |
5.7 |
3.5 |
3.5 |
39.2 |
32.3 |
17.0 |
29.3 |
|||||||||||||||||
POSITIVE CONTROL VALUES 2016 |
|
||||||||||||||||||||||||||||||||
Strain S9-Mix |
TA100 |
TA1535 |
TA102 |
WP2uvrA |
TA98 |
TA1537 |
WP2uvrA pKM101 |
WP2pKM101 |
|
||||||||||||||||||||||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
|||||||||||||||||
Values |
409 |
406 |
381 |
386 |
30 |
28 |
341 |
335 |
388 |
385 |
379 |
381 |
14 |
24 |
8 |
16 |
|
||||||||||||||||
Min |
221 |
284 |
84 |
92 |
897 |
629 |
107 |
102 |
100 |
96 |
95 |
101 |
445 |
574 |
1674 |
372 |
|
||||||||||||||||
Max |
2222 |
2863 |
2994 |
879 |
2326 |
2140 |
1611 |
637 |
449 |
4357 |
1413 |
639 |
1117 |
1855 |
2823 |
945 |
|
||||||||||||||||
Mean |
724 |
1264 |
854 |
240 |
1633 |
950 |
718 |
240 |
186 |
188 |
406 |
290 |
743 |
1271 |
2379 |
535 |
|
||||||||||||||||
SD |
320.4 |
562.9 |
664.9 |
62.1 |
564.5 |
382.7 |
338.6 |
98.2 |
49.8 |
230.8 |
227.0 |
92.7 |
214.6 |
326.5 |
426.2 |
143.3 |
|
||||||||||||||||
SD: Standard deviation
Min: Minimum value
Max: Maximum value
†: No. of mean values used to create dataset
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
In a reverse gene mutation assay in bacteria, performed according to the OECD Guideline 471 and in compliance with GLP, Salmonella typhimurium strains TA1535, TA1537, TA98 and TA100, and Escherichia coli strain WP2uvrA were exposed to the test item at the following concentrations:
- Experiment 1 - Pre incubation Method: 1.5, 5, 15, 50, 150, 500, 1500 and 5000 µg/plate, with and without S9-mix
- Experiment 2 - Pre-Incubation Method: 0.5, 1.5, 5, 15, 50, 150, 500 µg/plate, without S9-mix for TA100, TA1535 and TA 1537 and 5, 15, 50, 150, 500, 1500 and 5000 µg/plate for TA98 without S9. 15, 50, 150, 500, 1500and 5000 μg/plate with S9 for all strains.
Rat liver homogenate (10% liver S9 in standard co-factors) was used as a metabolizing system. Vehicle control, negative (untreated) and positive control groups were also included in mutagenicity tests.
The vehicle (acetone) 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.
In the first mutation test, the test item caused a visible reduction in the growth of the bacterial background lawns of all of the bacterial tester strains from 150 µg/plate (TA100, TA 1535 and TA 1537) and 1500 µg/plate (TA98 and WP2uvrA) in the absence of S9 mix and to two bacterial strains (TA100 and TA1535) at 5000 µg/plate in the presence S9-mix. In the second mutation test, the test item again induced a toxic response with weakened bacterial background lawns noted in the absence of S9-mix from 150 µg/plate (TA1535, TA100 and TA1537) and 1500 µg/plate (TA98 and WP2uvrA). In the presence of S9-mix, weakened bacterial background lawns were noted to three of the bacterial tester strains at 5000 µg/plate (TA100, TA1535 and WP2uvrA).
A test item precipitate (light and globular in appearance) was noted at 5000 µg/plate, this observation did not prevent the scoring of revertant colonies.
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 1 and Experiment 2 .
Under the test conditions, the test item is not considered as mutagenic in these bacterial systems.
Justification for classification or non-classification
Harmonized classification:
The test material has no harmonized classification for human health according to the Regulation (EC) No. 1272/2008.
Self-classification:
Based on the available data, no additional classification is proposed regarding germ cell mutagenicity according to the Regulation (EC) No. 1272/2008 (CLP)and to the Globally Harmonised System of classification and labelling of chemicals (GHS).
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