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EC number: 237-377-8 | CAS number: 13767-32-3
- 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
- Nanomaterial dustiness
- Nanomaterial porosity
- Nanomaterial pour density
- Nanomaterial photocatalytic activity
- Nanomaterial radical formation potential
- Nanomaterial catalytic activity
- Endpoint summary
- Stability
- Biodegradation
- Bioaccumulation
- 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
- Biological effects monitoring
- Biotransformation and kinetics
- Additional ecotoxological information
- Toxicological Summary
- Toxicokinetics, metabolism and distribution
- Acute Toxicity
- Irritation / corrosion
- Sensitisation
- Repeated dose toxicity
- 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
Key study: OECD 471. GLP study. It was concluded that the substance is not mutagenic in the Salmonella typhimurium test strains TA97a, TA98, TA100, TA102 and TA1535 up to 5000 μg/plate in the absence and presence of 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:
- 14 August 2018 - 23 August 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)
- 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
- Species / strain / cell type:
- S. typhimurium, other: TA97, TA98, TA100, TA102, TA1535
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9-Mix (rat liver S9)
- Test concentrations with justification for top dose:
- First experiment: 5000 μg/plate, 1500 μg/plate, 500 μg/plate, 150 μg/plate and 50 μg/plate.
Second experiment: 5000 μg/plate, 2500 μg/plate, 1250 μg/plate, 625 μg/plate, 313 μg/plate and 156 μg/plate. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: acetone
- Justification for choice of solvent/vehicle: In a non-GLP pre-test, the solubility of the test item was tested in demineralized water, di-methyl sulfoxide (DMSO), acetone and tetra hydrofurane. The solid test item was not sufficiently soluble in all solvents. Based on the results of the non-GLP pre-test, acetone was chosen as vehicle, because this vehicle showed a pipetable suspension it does not have any effects on the viability of the bacteria or the number of spontaneous revertants in the tested concentrations. - Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO, Demineralised water and Acetone
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- benzo(a)pyrene
- other: 4-Nitro-1,2-phenylene Diamine (20 µg in DMSO) - TA97a, TA98, TA102 (-S9) // 2-Amino-Anthracene (1 µg in DMSO) - TA97, TA100, TA102 and TA1535 (S9)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION:
First experiment: Plate incorporation method.
Second experiment: Pre-incubation method.
Per bacteria strain and concentration, three plates with and three plates without metabolic activation (-S9) were used. For the top agar 100 mL agar basis was melted in a microwave oven, 10 mL of the histidinebiotin-solution 0.5 mM was added, then the mixture was placed in the water bath at 43 ±1 °C. The following materials were gently vortexed in a test tube and poured onto the selective agar plates: 100 μL test suspension at each dose level, solvent (negative control) or reference
mutagen solution (positive control); 500 μL S9 mix; 100 μL bacteria suspension; 2000 μL overlay agar. In the second experiment, after the pre-incubation for 20 minutes, 2000 μL top agar was added. The plates were closed and left to solidify for a few minutes, then inverted and placed in the dark incubator at 37 ±1 °C for 48 h.
DURATION
- Preincubation period: 20 min (second experiment).
- Exposure duration: Incubation for 48 hours at 37 ± 1°C.
NUMBER OF REPLICATIONS: 3
EVALUATION:
The colonies were counted visually and the numbers were recorded. The mean values and standard deviations of each threefold determination was calculated as well as the increase factor f(l) of revertant induction (mean revertants divided by mean spontaneous revertants) of the test item suspensions and the positive controls. Additionally, the absolute number of revertants (mean revertants less mean spontaneous revertants) was given.
OTHER:
Genotype Confirmation:
Genotype confirmation is performed for each batch of lyophilized bacteria before stock cul-ture preparation.
Histidine requirement:
Each strain was streaked on a biotin and a histidine-biotin-plate, using a sterilized wire loop. The plates were incubated for 24 hours at 37 ±1 °C.
Ampicillin/Tetracycline-Resistance (pKM 101, pAQ1)
Each strain was streaked on an ampicillin agar plate and on an ampicillin-tetracycline agar plate. TA1535 was used as control strain, since it is not ampicillin resistant. The plates were incubated for 24 hours at 37 ±1 °C.
UV-sensitivity (uvrB):
Each strain was streaked on a plate, and one half of the plate covered with aluminium foil so that one half of each streak was protected against light. The plates for the strain TA97a, TA100 and TA102 were irradiated for 8 seconds, the plates for the strain TA 98 were irradi-ated for 10 seconds and the plates for the strain TA1535 were irradiated for 6 seconds with a germicidal lamp (254 nm, 30W). Keeping a distance of 33 cm for the strains TA97a, TA102 and TA1535. Keeping a distance of 66 cm for the following strains: TA98, TA100. Incubation for 24 hours at 37 ±1 °C followed.
Crystal violet sensitivity (deep rough/rfa):
For each strain, two plates were used. 0.1 mL of bacteria suspension were mixed with 2 mL Top-Agar and poured on nutrient agar. Sterile paper discs (Ø 9 mm), each soaked with 10 μL of crystal violet solution (0.1%) were placed into the middle of each plate, followed by incubation for 24 hours at 37 ±1°C.
Determination of Titre:
The titre was determined by dilution of the overnight culture using sodium chloride solution and placing 0.1 mL on maximal-soft agar. Incubation for 48 hours at 37 ±1 °C followed. It should give a density of 109 cells/mL (at the least), two replicates with and without metabolic activation.
Toxicity Control
Performed in experiment 1 only analogously to the titre control with the maximum dose of test item on maximal-soft agar, two replicates with and without metabolic activation, incubation for 48 hours at 37 ±1°C. - Evaluation criteria:
- A substance is considered to have mutagenic potential, if a reproducible increase of revertant colonies per plate exceeding an increase factor of 2 in at least one strain can be observed. A concentration-related increase over the range tested is also taken as a sign of mutagenic activity.
- Key result
- Species / strain:
- S. typhimurium, other: TA97, TA98, TA100, TA102, TA1535
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- no cytotoxicity
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Precipitation: In the first experiment, the test item showed precipitates on the plates in the two highest concentrations (5000 and 1500 μg/plate) in all bacteria strains. In the second experiment, the test item showed precipitates in the two highest concentrations (5000 and 2500 μg/plate) in all bacteria strains.
- Definition of acceptable cells for analysis: All strains met the criterion of at least 109 bacteria/mL, and no inconsistencies were found in the sterility control.
- Other confounding effects: Concentrations of the test item are stated as nominal concentrations, as suspensions had to be tested due to the poor solubility of the test item. As no complete dissolution was possible, undissolved particles were visible on the plates.
HISTORICAL CONTROL DATA (with ranges, means and standard deviation and confidence interval (e.g. 95%)
- Positive historical control data: All positive controls (diagnostic mutagens) showed mutagenic effects with and without metabolic activation and were within the historical control data ranges.
- Negative (solvent/vehicle) historical control data: Nearly all determined values for the spontaneous revertants of the negative controls were in the normal range of the test laboratory. - Conclusions:
- It was concluded that the substance is not mutagenic in the Salmonella typhimurium test strains TA97a, TA98, TA100, TA102 and TA1535 up to 5000 μg/plate in the absence and presence of metabolic activation under the experimental conditions in the present study.
- Executive summary:
An in-vitro bacterial reverse mutation test was performed according to the OECD Guideline 471 (GLP study). The test item was tested in the Salmonella typhimurium reverse mutation assay with five strains of Salmonella typhimurium (TA97a, TA98, TA100, TA102 and TA1535). The test was performed in two experiments up to concentrations of 5000 μg/plate (suspended in acetone) of in the presence and absence of metabolic activation. In the first experiment, the plate incorporation method was used. In the second experiment, the pre-incubation method was used. The test item showed precipitates on the plates at the two highest concentrations (5000 and 1500 μg/plate in the first experiment and 5000 μg/plate and 2500 μg/plate) in all bacteria strains. The bacterial background lawn was not reduced at any of the concentrations and no relevant decrease in the number of revertants was observed in all bacteria strains. The test item showed no signs of toxicity towards the bacteria strains in both the absence and presence of metabolic activation. The results of these experiments showed that the test item caused no increase in the number of revertants in all bacteria strains compared to the solvent control, in both the absence and presence of metabolic activation. The test item did not induce a dose-related increase in the number of revertants colonies in all strains, in the presence and absence of metabolic activation. Based on the results of this study it is concluded that the substance is not mutagenic in the Salmonella typhimurium strains TA97a, TA98, TA100, TA102 and TA1535 in the absence and presence of metabolic activation under the experimental conditions in this study.
Reference
Mean revertants first experiment:
Strain |
TA97a |
TA98 |
TA100 |
TA102 |
TA1535 |
||||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
Demin. water |
Mean |
82 |
88 |
38 |
42 |
102 |
118 |
355 |
331 |
10 |
12 |
sd |
1.0 |
4.5 |
1.2 |
1.7 |
11.6 |
20.6 |
148.7 |
30.0 |
1.2 |
3.0 |
|
DMSO |
Mean |
87 |
129 |
40 |
51 |
99 |
111 |
292 |
301 |
10 |
12 |
sd |
18.4 |
23.7 |
1.2 |
2.5 |
15.9 |
17.5 |
48.5 |
19.7 |
1.5 |
1.7 |
|
Acetone |
Mean |
130 |
120 |
47 |
48 |
95 |
123 |
471 |
293 |
12 |
11 |
sd |
22.4 |
10.7 |
5.1 |
2.9 |
11.0 |
14.2 |
128.0 |
20.1 |
2.5 |
2.6 |
|
Positive |
Mean |
613 |
580 |
131 |
295 |
417 |
1001 |
1467 |
1232 |
437 |
146 |
sd |
68.0 |
181.6 |
11.6 |
34.0 |
43.9 |
0.0 |
95.4 |
284.4 |
52.1 |
40.8 |
|
f(I) |
7.05 |
4.50 |
3.28 |
5.78 |
4.09 |
9.02 |
5.02 |
4.09 |
43.70 |
12.17 |
|
5000 µg/plate |
Mean |
97 |
102 |
43 |
40 |
113 |
115 |
384 |
397 |
12 |
11 |
sd |
19.1 |
17.1 |
5.7 |
4.2 |
12.9 |
13.3 |
26.2 |
41.1 |
3.0 |
0.0 |
|
f(I) |
0.75 |
0.85 |
0.91 |
0.83 |
1.19 |
0.93 |
0.82 |
1.35 |
1.00 |
1.00 |
|
1500 µg/plate |
Mean |
82 |
83 |
50 |
39 |
87 |
97 |
393 |
347 |
11 |
11 |
sd |
11.0 |
10.0 |
2.6 |
5.3 |
6.6 |
6.1 |
4.6 |
28.4 |
1.5 |
0.6 |
|
f(I) |
0.63 |
0.69 |
1.06 |
0.81 |
0.92 |
0.79 |
0.83 |
1.18 |
0.92 |
1.00 |
|
500 µg/plate |
Mean |
93 |
106 |
45 |
46 |
89 |
113 |
309 |
307 |
12 |
13 |
sd |
1.7 |
31.0 |
3.6 |
5.0 |
7.0 |
7.0 |
23.4 |
23.4 |
2.1 |
1.2 |
|
f(I) |
0.72 |
0.88 |
0.96 |
0.96 |
0.94 |
0.92 |
0.66 |
1.05 |
1.00 |
1.18 |
|
150 µg/plate |
Mean |
89 |
77 |
43 |
42 |
107 |
115 |
292 |
295 |
12 |
11 |
sd |
7.0 |
3.2 |
11.1 |
9.0 |
11.0 |
14.0 |
43.3 |
4.6 |
2.0 |
1.5 |
|
f(I) |
0.68 |
0.64 |
0.91 |
0.88 |
1.13 |
0.93 |
0.62 |
1.01 |
1.00 |
1.00 |
|
50 µg/plate |
Mean |
94 |
84 |
50 |
46 |
105 |
117 |
287 |
283 |
13 |
13 |
sd |
17.0 |
10.1 |
4.6 |
6.1 |
1.2 |
11.0 |
28.9 |
15.1 |
3.2 |
2.0 |
|
f(I) |
0.72 |
0.70 |
1.06 |
0.96 |
1.11 |
0.95 |
0.61 |
0.97 |
1.08 |
1.18 |
f(I) = increase factor
1001 colonies per plate means the bacteria growth was too strong for counting.
Mean revertants second experiment:
Strain |
TA97a |
TA98 |
TA100 |
TA102 |
TA1535 |
||||||
Induction |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
-S9 |
+S9 |
|
Demin. water |
Mean |
95 |
91 |
45 |
48 |
101 |
114 |
245 |
260 |
13 |
11 |
sd |
12.1 |
15.1 |
4.7 |
4.2 |
4.5 |
17.1 |
28.4 |
27.7 |
2.0 |
1.5 |
|
DMSO |
Mean |
116 |
99 |
47 |
48 |
81 |
101 |
232 |
236 |
12 |
12 |
sd |
27.8 |
25.9 |
7.1 |
3.6 |
5.5 |
2.3 |
26.2 |
28.0 |
1.2 |
2.9 |
|
Acetone |
Mean |
99 |
113 |
45 |
51 |
105 |
104 |
237 |
271 |
13 |
11 |
sd |
10.6 |
27.0 |
3.8 |
3.0 |
28.6 |
15.0 |
2.3 |
49.4 |
2.9 |
2.1 |
|
Positive |
Mean |
635 |
627 |
181 |
177 |
643 |
895 |
1432 |
1448 |
232 |
90 |
sd |
156.0 |
60.7 |
10.1 |
12.2 |
34.0 |
183.0 |
44.5 |
28.8 |
58.1 |
10.2 |
|
f(I) |
5.47 |
6.33 |
3.85 |
3.69 |
6.37 |
8.86 |
6.17 |
6.14 |
17.85 |
7.50 |
|
5000 µg/plate |
Mean |
89 |
122 |
48 |
50 |
106 |
107 |
257 |
249 |
11 |
10 |
sd |
12.9 |
7.8 |
6.4 |
2.1 |
14.0 |
9.9 |
50.0 |
47.4 |
3.2 |
0.6 |
|
f(I) |
0.90 |
1.08 |
1.07 |
0.98 |
1.01 |
1.03 |
1.08 |
0.92 |
0.85 |
0.91 |
|
2500 µg/plate |
Mean |
98 |
146 |
50 |
48 |
104 |
107 |
215 |
211 |
10 |
12 |
sd |
10.4 |
2.1 |
3.8 |
5.8 |
2.0 |
4.2 |
8.3 |
18.9 |
2.0 |
1.5 |
|
f(I) |
0.99 |
1.29 |
1.11 |
0.94 |
0.99 |
1.03 |
0.91 |
0.78 |
0.77 |
1.09 |
|
1250 µg/plate |
Mean |
84 |
147 |
50 |
46 |
97 |
124 |
227 |
208 |
10 |
10 |
sd |
1.0 |
27.9 |
3.1 |
2.5 |
6.4 |
2.5 |
25.7 |
8.0 |
0.6 |
2.5 |
|
f(I) |
0.85 |
1.30 |
1.11 |
0.90 |
0.92 |
1.19 |
0.96 |
0.77 |
0.77 |
0.91 |
|
625 µg/plate |
Mean |
92 |
125 |
49 |
50 |
99 |
113 |
228 |
261 |
10 |
11 |
sd |
6.4 |
8.0 |
6.8 |
5.3 |
3.1 |
16.2 |
48.7 |
22.0 |
2.0 |
1.7 |
|
f(I) |
0.93 |
1.11 |
1.09 |
0.98 |
0.94 |
1.09 |
0.96 |
0.96 |
0.77 |
1.00 |
|
312 µg/plate |
Mean |
87 |
118 |
45 |
48 |
108 |
123 |
223 |
225 |
9 |
11 |
sd |
4.9 |
30.4 |
5.9 |
6.2 |
11.5 |
30.3 |
24.4 |
16.2 |
2.6 |
2.3 |
|
f(I) |
0.88 |
1.04 |
1.00 |
0.94 |
1.03 |
1.18 |
0.94 |
0.83 |
0.69 |
1.00 |
|
156 µg/plate |
Mean |
83 |
120 |
53 |
50 |
101 |
114 |
225 |
236 |
9 |
10 |
sd |
2.6 |
14.0 |
1.5 |
4.2 |
8.0 |
11.7 |
54.6 |
50.1 |
1.7 |
1.0 |
|
f(I) |
0.84 |
1.06 |
1.18 |
0.98 |
0.96 |
1.10 |
0.95 |
0.87 |
0.69 |
0.91 |
f(I) = increase factor
Historical data of espontaneous revertants:
Strain |
|
TA97a |
TA98 |
TA100 |
TA102 |
TA1535 |
|||||
Induction |
|
- S9 |
+ S9 |
- S9 |
+ S9 |
- S9 |
+ S9 |
- S9 |
+ S9 |
- S9 |
+ S9 |
Demin. water |
Mean |
88 |
95 |
24 |
26 |
93 |
98 |
283 |
302 |
18 |
17 |
Min |
60 |
63 |
6 |
8 |
51 |
64 |
85 |
67 |
6 |
7 |
|
Max |
144 |
138 |
52 |
53 |
147 |
141 |
445 |
587 |
36 |
40 |
|
SD |
17 |
17 |
13 |
12 |
15 |
15 |
60 |
72 |
6 |
6 |
|
Exp 1 |
82 |
88 |
38 |
42 |
102 |
118 |
355 |
331 |
10 |
12 |
|
Exp 2 |
95 |
91 |
45 |
48 |
101 |
114 |
245 |
260 |
13 |
11 |
|
DMSO |
Mean |
89 |
98 |
24 |
25 |
90 |
93 |
285 |
297 |
17 |
17 |
Min |
58 |
67 |
7 |
8 |
44 |
62 |
79 |
80 |
8 |
6 |
|
Max |
139 |
144 |
50 |
50 |
138 |
199 |
465 |
499 |
35 |
37 |
|
SD |
17 |
16 |
13 |
12 |
15 |
17 |
61 |
63 |
6 |
6 |
|
Exp 1 |
87 |
129 |
40 |
51 |
99 |
111 |
292 |
301 |
10 |
12 |
|
Exp 2 |
116 |
99 |
47 |
48 |
81 |
101 |
232 |
236 |
12 |
12 |
|
Acetone |
Mean |
70 |
107 |
21 |
24 |
76 |
84 |
330 |
336 |
17 |
17 |
Min |
34 |
61 |
6 |
11 |
52 |
54 |
213 |
223 |
10 |
10 |
|
Max |
88 |
142 |
52 |
51 |
147 |
140 |
525 |
485 |
36 |
40 |
|
SD |
16 |
25 |
14 |
13 |
22 |
21 |
79 |
77 |
6 |
6 |
|
Exp 1 |
130 |
120 |
47 |
48 |
95 |
123 |
471 |
293 |
12 |
11 |
|
Exp 2 |
99 |
113 |
45 |
51 |
105 |
104 |
237 |
271 |
13 |
11 |
|
Positive Controls |
Mean |
539 |
537 |
403 |
138 |
507 |
797 |
1104 |
1205 |
266 |
141 |
Min |
264 |
228 |
77 |
39 |
220 |
273 |
491 |
408 |
55 |
45 |
|
Max |
1165 |
1181 |
1001 |
487 |
1256 |
1912 |
2331 |
6083 |
515 |
712 |
|
SD |
166 |
176 |
171 |
105 |
187 |
285 |
397 |
539 |
85 |
84 |
|
Exp 1 |
613 |
580 |
131 |
295 |
417 |
1001 |
1467 |
1232 |
437 |
146 |
|
Exp 2 |
635 |
627 |
181 |
177 |
643 |
895 |
1432 |
1448 |
232 |
90 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
Justification for classification or non-classification
Based on available data, the substance is not classified for mutagenicity according to the CLP Regulation (EC) no. 1272/2008.
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.