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EC number: 810-766-5 | CAS number: 1802886-42-5
- Life Cycle description
- Uses advised against
- Endpoint summary
- Appearance / physical state / colour
- Melting point / freezing point
- Boiling point
- Density
- Particle size distribution (Granulometry)
- Vapour pressure
- Partition coefficient
- Water solubility
- Solubility in organic solvents / fat solubility
- Surface tension
- Flash point
- Auto flammability
- Flammability
- Explosiveness
- Oxidising properties
- Oxidation reduction potential
- Stability in organic solvents and identity of relevant degradation products
- Storage stability and reactivity towards container material
- Stability: thermal, sunlight, metals
- pH
- Dissociation constant
- Viscosity
- Additional physico-chemical information
- Additional physico-chemical properties of nanomaterials
- Nanomaterial agglomeration / aggregation
- Nanomaterial crystalline phase
- Nanomaterial crystallite and grain size
- Nanomaterial aspect ratio / shape
- Nanomaterial specific surface area
- Nanomaterial Zeta potential
- Nanomaterial surface chemistry
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- Endpoint summary
- Stability
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- Transport and distribution
- Environmental data
- Additional information on environmental fate and behaviour
- Ecotoxicological Summary
- Aquatic toxicity
- Endpoint summary
- Short-term toxicity to fish
- Long-term toxicity to fish
- Short-term toxicity to aquatic invertebrates
- Long-term toxicity to aquatic invertebrates
- Toxicity to aquatic algae and cyanobacteria
- Toxicity to aquatic plants other than algae
- Toxicity to microorganisms
- Endocrine disrupter testing in aquatic vertebrates – in vivo
- Toxicity to other aquatic organisms
- Sediment toxicity
- Terrestrial toxicity
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- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
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- Genetic toxicity
- Carcinogenicity
- Toxicity to reproduction
- Specific investigations
- Exposure related observations in humans
- Toxic effects on livestock and pets
- Additional toxicological data
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Based on an bacterial reverse mutation test according to OECD guideline 471 the test substance was considered to be non-mutagenic with and without metabolic activation.
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:
- 18 November 2015 - 07 March 2018
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 1997
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
- Version / remarks:
- 2008
- Deviations:
- no
- Qualifier:
- according to guideline
- Guideline:
- EPA OPPTS 870.5100 - Bacterial Reverse Mutation Test (August 1998)
- Version / remarks:
- 1998
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and E. coli WP2
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 fraction of Phenobarbital (PB) and β-naphthoflavone (BNF) induced rat liver
- Test concentrations with justification for top dose:
- Concentrations: 5, 16, 50, 160, 500 and 1000 μg/plate (-S9 mix), 16, 50, 160, 500, 1000 and 1600 μg/plate (+S9 mix)
Justification for top dose: Based on a concentration range finding test. In this test strong cytotoxic effect of the test item (absent background lawn and absent revertant growth) was obtained at the concentrations of 5000 and 1600 μg/plate, without metabolic activation (-S9 Mix) and at 5000 μg/plate, with addition of metabolic activation (+S9 Mix). - Vehicle / solvent:
- - Vehicle used: DMSO
- Justification for choice of solvent/vehicle: Based on a solubility test. - Untreated negative controls:
- no
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- 9-aminoacridine
- sodium azide
- methylmethanesulfonate
- other: 2-aminoanthracene and 4-nitro-1,2-phenylene-diamine
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation) and preincubation
DURATION
- Preincubation period: 20 minutes, 37 °C
- Exposure duration: 48 hours
NUMBER OF REPLICATIONS: 3 (two independent experiments)
DETERMINATION OF CYTOTOXICITY
- Method: counting numbers of revertants - Evaluation criteria:
- A test item is considered mutagenic if:
- a dose-related increase in the number of revertants occurs and/or;
- a reproducible biologically relevant positive response for at least one of the dose groups occurs in at least one strain with or without metabolic activation.
An increase is considered biologically relevant if:
- in strain TA 100 the number of reversions is at least twice as high as the reversion rate of the vehicle control
- in strain TA 98, TA 1535, TA 1537 and Escherichia coli WP2 uvrA the number of reversions is at least three times higher than the reversion rate of the vehicle control. - Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 160 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1535
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 1000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 160 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 1537
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 1000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 160 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 98
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 1000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 500 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- S. typhimurium TA 100
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 1000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 500 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Key result
- Species / strain:
- E. coli WP2 uvr A
- Metabolic activation:
- with
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- at ≥ 1600 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- not examined
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: No precipitation of the test item was observed .
RANGE-FINDING/SCREENING STUDIES: The concentration range investigated in the main tests was based on two preliminary range finding tests. - Conclusions:
- The test substance was considered to be non-mutagenic with and without metabolic activation in bacteria.
- Executive summary:
The test item was examined for its mutagenic activity in two series of in vitro microbial assays employing Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537, and Escherichia coli WP2 uvrA as indicator organisms. The plate incorporation test with and without addition of liver S9 mix from phenobarbital/p-naphthoflavone-pretreated rats was used. In this study, two experimental series were performed. In the experiments with S9 mix, 10% and 20% S9 in the S9 mix were used. Treatments of all tester strains were performed using test item formulations prepared in DMSO in the absence and in the presence of S9 mix, using final concentrations between 5 and 1000 µg/plate (-S9) and between 16 and 1600 µg/plate (+S9), plus vehicle and positive controls.The concentrations, including the controls, were tested in triplicates. In the performed experiments positive and negative (vehicle) controls were run concurrently. For the analysis of the test item concentrations, representative samples from the stock solutions (with a nominal concentration of 32 mg/mL) and vehicle control were taken and analysed before each main experiment.
In the performed experiments all of the validity criteria, regarding the investigated strains, negative (vehicle) and positive controls, S9 activity and number of investigated analysable concentration levels were fulfilled. No biological relevant increases were observed in revertant colony numbers of any of the five test strains following treatment with the test item at any concentration level, either in the presence or absence of metabolic activation (S9 Mix) in the performed experiments. Sporadic increases in revertant colony numbers compared to the vehicle control values within the actual historical control data ranges were observed in both independently performed main experiments. However, there was no tendency of higher mutation rates with increasing concentrations beyond the generally acknowledged border of biological relevance in the performed experiments. In the performed experiments inhibitory effects of the test item (absent revertants, decreased number of revertant colony numbers and/or affected background lawn development) were observed in all examined strains. The 160 μg/plate was found to be the lowest cytotoxic concentration, observed in confirmatory mutation test in the case of Salmonella typhimurium TA98, TA1535 and TA1537 strains, in the absence of exogenous metabolic activation (-S9 Mix). No precipitation of the test item was observed on the plates in the examined bacterial strains at any examined concentration level (±S9 Mix) throughout the study.
The reported data of this mutagenicity assay showed, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution in the genome of the tester strains used. Therefore, the test item is considered non-mutagenicin this bacterial reverse mutation assay.
Reference
Analytical Measurment:
To verify the test item concentrations samples from the stock solution and vehicle control will be taken and the test item content was determined with a previously validated analytical method. For the analysis of the test item concentrations, representative samples from the stock solutions (with a concentration of 32 mg/mL) and vehicle control were taken and analysed before each main experiment. The two main components of test item were used for the quantitation of the test item. The measured concentrations of components mentioned above varied between 92 % and 102 % in the samples in comparison to the nominal values.
Table 1: Summary Table of the Results of the Initial Mutation Test
Initial Mutation Test (Plate Incorporation Test) |
||||||||||||||||||||
Concentrations (mg/plate) |
Salmonella typhimuriumtester strains |
Escherichia coli |
||||||||||||||||||
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
WP2uvrA |
||||||||||||||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|||||||||||
Mean values of revertants per plate Mutation rate (MR) |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Untreated Control |
19.3 |
1.07 |
21.3 |
0.98 |
82.0 |
1.21 |
91.3 |
1.02 |
9.3 |
0.88 |
9.0 |
0.96 |
4.0 |
0.67 |
5.3 |
0.84 |
33.0 |
1.03 |
42.7 |
1.24 |
DMSO Control (100 µL) |
17.7 |
1.00 |
24.0 |
1.00 |
– |
– |
80.7 |
1.00 |
– |
– |
8.7 |
1.00 |
3.7 |
1.00 |
4.7 |
1.00 |
– |
– |
33.0 |
1.00 |
Ultrapure Water Control |
– |
– |
– |
– |
79.7 |
1.00 |
– |
– |
9.7 |
1.00 |
– |
– |
– |
– |
– |
– |
26.0 |
1.00 |
– |
– |
DMSO Control (50 µL) |
18.0 |
1.00 |
21.7 |
1.00 |
68.0 |
1.00 |
89.3 |
1.00 |
10.7 |
1.00 |
9.3 |
1.00 |
6.0 |
1.00 |
6.3 |
1.00 |
32.0 |
1.00 |
34.3 |
1.00 |
1600 |
– |
– |
3.7 |
0.17 |
– |
– |
2.3 |
0.03 |
– |
– |
0.0 |
0.00 |
– |
– |
0.0 |
0.00 |
– |
– |
16.7 |
0.49 |
1000 |
0.0 |
0.00 |
18.3 |
0.85 |
0.0 |
0.00 |
117.0 |
1.31 |
0.0 |
0.00 |
15.3 |
1.64 |
0.0 |
0.00 |
4.3 |
0.68 |
13.0 |
0.41 |
36.0 |
1.05 |
500 |
6.0 |
0.33 |
38.3 |
1.77 |
72.7 |
1.07 |
138.7 |
1.55 |
4.3 |
0.41 |
19.7 |
2.11 |
0.0 |
0.00 |
5.7 |
0.89 |
33.3 |
1.04 |
40.0 |
1.17 |
160 |
20.0 |
1.11 |
34.7 |
1.60 |
113.3 |
1.67 |
127.3 |
1.43 |
11.3 |
1.06 |
16.3 |
1.75 |
4.3 |
0.72 |
8.3 |
1.32 |
35.7 |
1.11 |
33.0 |
0.96 |
50 |
20.3 |
1.13 |
36.3 |
1.68 |
89.0 |
1.31 |
101.0 |
1.13 |
9.0 |
0.84 |
13.7 |
1.46 |
7.3 |
1.22 |
7.0 |
1.11 |
35.7 |
1.11 |
37.7 |
1.10 |
16 |
14.3 |
0.80 |
26.7 |
1.23 |
86.7 |
1.27 |
101.3 |
1.13 |
9.0 |
0.84 |
11.0 |
1.18 |
5.3 |
0.89 |
8.0 |
1.26 |
27.0 |
0.84 |
39.7 |
1.16 |
5 |
17.3 |
0.96 |
– |
– |
79.7 |
1.17 |
– |
– |
9.3 |
0.88 |
– |
– |
7.3 |
1.22 |
– |
– |
34.3 |
1.07 |
– |
– |
NPD (4mg) |
481.3 |
27.25 |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
SAZ (2mg) |
– |
– |
– |
– |
796.7 |
10.00 |
– |
– |
1093.3 |
113.10 |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
9AA (50mg) |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
581.3 |
158.55 |
– |
– |
– |
– |
– |
– |
MMS (2mL) |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
562.7 |
21.64 |
– |
– |
2AA (2mg) |
– |
– |
1290.7 |
53.78 |
– |
– |
1629.3 |
20.20 |
– |
– |
194.3 |
22.42 |
– |
– |
123.0 |
26.36 |
– |
– |
– |
– |
2AA (50mg) |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
215.3 |
6.53 |
MR: Mutation Rate; NPD: 4-Nitro-1,2-phenylenediamine; SAZ: Sodium azide; 9AA: 9-Aminoacridine; MMS: Methyl methanesulfonate; 2AA: 2-aminoanthracene
Remarks: DMSO (50 µL) was applied as vehicle of the test item and DMSO (100 µL) was applied as vehicle of the positive control substances NPD, 9AA and 2AA. The ultrapure water (100 µL) was applied as vehicle of the positive control substances MMS and SAZ. The mutation rate of the test item and the untreated control refers to the DMSO sample (50 µL); the mutation rate of the NPD, 9AA and 2AA refers to the DMSO sample (100 µL). The mutation rate of MMS and SAZ refers to ultrapure water (100 µL).
Table 2: Summary Table of the Results of the Confirmatory Mutation Test
Confirmatory Mutation Test (Pre-Incubation Test) |
||||||||||||||||||||||
Concentrations (mg/plate) |
Salmonella typhimuriumtester strains |
Escherichia coli |
||||||||||||||||||||
TA 98 |
TA 100 |
TA 1535 |
TA 1537 |
WP2uvrA |
||||||||||||||||||
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|||||||||||||
Mean values of revertants per plate Mutation rate (MR) |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
Mean |
MR |
||
Untreated Control |
21.3 |
0.93 |
26.7 |
1.10 |
87.3 |
1.12 |
100.0 |
1.12 |
10.3 |
0.76 |
11.7 |
1.09 |
7.3 |
1.05 |
6.7 |
1.18 |
27.3 |
0.92 |
32.7 |
0.96 |
||
DMSO Control (100 µL) |
16.3 |
1.00 |
23.3 |
1.00 |
– |
– |
73.0 |
1.00 |
– |
– |
9.7 |
1.00 |
6.7 |
1.00 |
5.3 |
1.00 |
– |
– |
33.0 |
1.00 |
||
Ultrapure Water Control |
– |
– |
– |
– |
77.3 |
1.00 |
– |
– |
9.3 |
1.00 |
– |
– |
– |
– |
– |
– |
31.0 |
1.00 |
– |
– |
||
DMSO Control (50 µL) |
23.0 |
1.00 |
24.3 |
1.00 |
78.0 |
1.00 |
89.3 |
1.00 |
13.7 |
1.00 |
10.7 |
1.00 |
7.0 |
1.00 |
5.7 |
1.00 |
29.7 |
1.00 |
34.0 |
1.00 |
||
1600 |
– |
– |
0.0 |
0.00 |
– |
– |
0.0 |
1.00 |
– |
– |
0.0 |
0.00 |
– |
– |
0.0 |
0.00 |
– |
– |
10.3 |
0.30 |
||
1000 |
0.0 |
0.00 |
4.0 |
0.16 |
0.0 |
0.00 |
36.7 |
0.00 |
0.0 |
0.00 |
8.0 |
0.75 |
– |
– |
0.0 |
0.00 |
4.0 |
0.13 |
26.7 |
0.78 |
||
500 |
1.7 |
0.07 |
31.7 |
1.30 |
6.0 |
0.08 |
117.3 |
0.41 |
2.7 |
0.20 |
13.7 |
1.28 |
0.0 |
0.00 |
6.7 |
1.18 |
11.3 |
0.38 |
35.7 |
1.05 |
||
300 |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
1.3 |
0.19 |
– |
– |
– |
– |
– |
– |
||
160 |
11.0 |
0.48 |
32.3 |
1.33 |
70.7 |
0.91 |
112.0 |
1.25 |
7.7 |
0.56 |
10.3 |
0.97 |
2.7 |
0.38 |
8.3 |
1.47 |
40.3 |
1.36 |
38.0 |
1.12 |
||
50 |
28.7 |
1.25 |
31.7 |
1.30 |
88.7 |
1.14 |
101.7 |
1.14 |
11.0 |
0.80 |
13.0 |
1.22 |
7.0 |
1.00 |
7.3 |
1.29 |
29.7 |
1.00 |
35.3 |
1.04 |
||
16 |
24.0 |
1.04 |
31.7 |
1.30 |
95.3 |
1.22 |
112.3 |
1.26 |
11.7 |
0.85 |
14.0 |
1.31 |
6.7 |
0.95 |
8.0 |
1.41 |
32.3 |
1.09 |
32.0 |
0.94 |
||
5 |
27.7 |
1.20 |
23.7 |
0.97 |
84.0 |
1.08 |
130.7 |
1.46 |
11.3 |
0.83 |
13.3 |
1.25 |
8.3 |
1.19 |
6.0 |
1.06 |
27.0 |
0.91 |
40.3 |
1.19 |
||
1.6 |
23.7 |
1.03 |
– |
– |
97.3 |
1.25 |
– |
– |
13.3 |
0.98 |
– |
– |
7.7 |
1.10 |
– |
– |
28.7 |
0.97 |
– |
– |
||
NPD (4mg) |
452.0 |
27.67 |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
||
SAZ (2mg) |
– |
– |
– |
– |
1034.7 |
13.38 |
– |
– |
840.0 |
90.00 |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
||
9AA (50mg) |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
445.3 |
66.80 |
– |
– |
– |
– |
– |
– |
||
MMS (2mL) |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
978.7 |
31.57 |
– |
– |
||
2AA (2mg) |
– |
– |
965.3 |
41.37 |
– |
– |
1056.0 |
14.47 |
– |
– |
117.3 |
12.14 |
– |
– |
90.0 |
16.88 |
– |
– |
– |
– |
||
2AA (50mg) |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
– |
213.3 |
6.46 |
||
MR: Mutation Rate; NPD:4-Nitro-1,2-phenylenediamine; SAZ: Sodium azide; 9AA: 9-Aminoacridine; MMS: Methyl methanesulfonate; 2AA: 2-aminoanthracene
Remarks: DMSO (50 µL) was applied as vehicle of the test item and DMSO (100 µL) was applied as vehicle of the positive control substances NPD, 9AA and 2AA. The ultrapure water (100 µL) was applied as vehicle of the positive control substances MMS and SAZ. The mutation rate of the test item and the untreated control refers to the DMSO sample (50 µL); the mutation rate of the NPD, 9AA and 2AA refers to the DMSO sample (100 µL). The mutation rate of MMS and SAZ refers to ultrapure water (100 µL).
Table 3: Historical Control Values
|
Bacterial strains |
||||||
Historical control data of untreated control |
‑S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
Average |
19.7 |
94.6 |
10.9 |
8.9 |
25.2 |
||
SD |
1.7 |
2.4 |
0.4 |
1.3 |
5.2 |
||
Minimum |
8 |
67 |
4 |
3 |
11 |
||
Maximum |
40 |
133 |
21 |
20 |
52 |
||
+S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
|
Average |
24.5 |
112.9 |
11.0 |
9.2 |
31.7 |
||
SD |
1.4 |
6.1 |
0.5 |
1.2 |
6.4 |
||
Minimum |
11 |
74 |
3 |
3 |
13 |
||
Maximum |
43 |
159 |
20 |
20 |
60 |
||
|
Bacterial strains |
||||||
Historical control data of DMSO control |
‑S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
Average |
18.1 |
87.0 |
10.8 |
8.5 |
24.6 |
||
SD |
1.1 |
3.6 |
0.6 |
1.3 |
3.3 |
||
Minimum |
9 |
58 |
4 |
3 |
10 |
||
Maximum |
36 |
131 |
23 |
20 |
54 |
||
+S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
|
Average |
23.0 |
102.4 |
11.0 |
9.2 |
31.4 |
||
SD |
0.8 |
7.7 |
0.4 |
1.3 |
5.8 |
||
Minimum |
11 |
69 |
3 |
3 |
12 |
||
Maximum |
42 |
148 |
23 |
21 |
59 |
||
|
Bacterial strains |
||||||
Historical control data of Water control |
‑S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
Average |
19.5 |
92.8 |
11.3 |
9.1 |
26.6 |
||
SD |
1.4 |
5.0 |
0.5 |
1.8 |
6.6 |
||
Minimum |
12 |
62 |
4 |
3 |
10 |
||
Maximum |
30 |
139 |
22 |
18 |
52 |
||
+S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
|
Average |
24.4 |
109.9 |
11.2 |
9.5 |
34.0 |
||
SD |
1.2 |
6.7 |
0.8 |
1.9 |
6.1 |
||
Minimum |
13 |
83 |
5 |
4 |
16 |
||
Maximum |
37 |
149 |
18 |
18 |
63 |
||
|
Bacterial strains |
||||||
Historical control data of positive controls |
‑S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
Average |
285.9 |
1214.0 |
1024.3 |
594.4 |
858.4 |
||
SD |
25.5 |
80.0 |
120.3 |
36.4 |
164.7 |
||
Minimum |
152 |
609 |
407 |
136 |
330 |
||
Maximum |
598 |
2272 |
2597 |
2048 |
1760 |
||
+S9 |
|
TA98 |
TA100 |
TA1535 |
TA1537 |
E. coli |
|
Average |
1395.7 |
1727.4 |
160.7 |
145.8 |
204.9 |
||
SD |
261.4 |
244.1 |
30.0 |
18.9 |
3.9 |
||
Minimum |
286 |
712 |
91 |
70 |
133 |
||
Maximum |
3211 |
3435 |
328 |
315 |
367 |
Abbreviations: TA98, TA100, TA1535, TA1537: Salmonella typhimurium TA98, TA100, TA1535, TA1537; E. coli: Escherichia coliWP2uvrA, SD: Standard deviation
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Endpoint conclusion
- Endpoint conclusion:
- no study available
Additional information
The test item was examined for its mutagenic activity in two series of in vitro microbial assays employing Salmonella typhimurium TA 98, TA 100, TA 1535 and TA 1537, and Escherichia coli WP2 uvrA as indicator organisms. The plate incorporation test with and without addition of liver S9 mix from phenobarbital/p-naphthoflavone-pretreated rats was used. In this study, two experimental series were performed. In the experiments with S9 mix, 10% and 20% S9 in the S9 mix were used. Treatments of all tester strains were performed using test item formulations prepared in DMSO in the absence and in the presence of S9 mix, using final concentrations between 5 and 1000 µg/plate (-S9) and between 16 and 1600 µg/plate (+S9), plus vehicle and positive controls. The concentrations, including the controls, were tested in triplicates. In the performed experiments positive and negative (vehicle) controls were run concurrently. For the analysis of the test item concentrations, representative samples from the stock solutions (with a nominal concentration of 32 mg/mL) and vehicle control were taken and analysed before each main experiment.
In the performed experiments all of the validity criteria, regarding the investigated strains, negative (vehicle) and positive controls, S9 activity and number of investigated analysable concentration levels were fulfilled. No biological relevant increases were observed in revertant colony numbers of any of the five test strains following treatment with the test item at any concentration level, either in the presence or absence of metabolic activation (S9 Mix) in the performed experiments. Sporadic increases in revertant colony numbers compared to the vehicle control values within the actual historical control data ranges were observed in both independently performed main experiments. However, there was no tendency of higher mutation rates with increasing concentrations beyond the generally acknowledged border of biological relevance in the performed experiments. In the performed experiments inhibitory effects of the test item (absent revertants, decreased number of revertant colony numbers and/or affected background lawn development) were observed in all examined strains. The 160μg/plate was found to be the lowest cytotoxic concentration, observed in confirmatory mutation test in the case of Salmonella typhimurium TA98, TA1535 and TA1537 strains, in the absence of exogenous metabolic activation (-S9 Mix). No precipitation of the test item was observed on the plates in the examined bacterial strains at any examined concentration level (±S9 Mix) throughout the study.
The reported data showed, that under the experimental conditions reported, the test item did not induce gene mutations by frameshift or base-pair substitution in the genome of the tester strains used. Therefore, the test item is considered non-mutagenic in this bacterial reverse mutation assay.
Justification for classification or non-classification
Classification, Labelling, and Packaging Regulation (EC) No 1272/2008
The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. The results indicate that the substance is non-mutagenic. Based on available data the test item will not be classified for genetic toxicity according to Regulation (EC) No 1272/2008, as amended for the fifteenth time in Regulation (EU) 2020/1182.
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