Registration Dossier
Registration Dossier
Data platform availability banner - registered substances factsheets
Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.
The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.
Diss Factsheets
Use of this information is subject to copyright laws and may require the permission of the owner of the information, as described in the ECHA Legal Notice.
EC number: 274-157-0 | CAS number: 69851-61-2
- 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
Genetic toxicity: in vitro
Administrative data
- Endpoint:
- in vitro gene mutation study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- guideline study
Data source
Reference
- Reference Type:
- study report
- Title:
- Unnamed
- Year:
- 2 018
- Report date:
- 2018
Materials and methods
Test guideline
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- yes
- Remarks:
- To clarify equivocal results an independent repetition experiment (without metabolic activation) with closer spaced concentrations and double cell cultures was performed on sponsor’s request.
- GLP compliance:
- yes
- Type of assay:
- other: HPRT assay in Chinese Hamster V79 cells
Test material
- Reference substance name:
- N,N'-propane-1,3-diylbis[3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionamide]
- EC Number:
- 274-157-0
- EC Name:
- N,N'-propane-1,3-diylbis[3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionamide]
- Cas Number:
- 69851-61-2
- Molecular formula:
- C37H58N2O4
- IUPAC Name:
- N,N'-propane-1,3-diylbis[3-(3,5-di-tert-butyl-4-hydroxyphenyl)propanamide]
- Test material form:
- solid: particulate/powder
- Details on test material:
- - Purity: 98.4%
Constituent 1
Method
- Target gene:
- HPRT locus using V79 cells of the Chinese Hamster
Species / strain
- Species / strain / cell type:
- Chinese hamster lung fibroblasts (V79)
- Details on mammalian cell type (if applicable):
- CELLS USED
- Source of cells: Eurofins BioPharma Product Testing Munich GmbH stock cultures
- Cell cycle length, doubling time or proliferation index:12 - 14 h doubling time; high cloning efficiency of untreated cells, usually more than 50%
- Methods for maintenance in cell culture if applicable: The V79 cells (ATCC, CCL-93) were stored over liquid nitrogen (vapour phase) in the cell bank at the testing facility.
MEDIA USED
- Type and identity of media including CO2 concentration if applicable: stored over liquid nitrogen (vapour phase); minimal essential medium (MEM)
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes (via PCR)
- Periodically checked for karyotype stability: yes
- Periodically 'cleansed' against high spontaneous background: yes
- Metabolic activation:
- with and without
- Metabolic activation system:
- rat liver (S9)
- Test concentrations with justification for top dose:
- Toxicity test: 1, 2.5, 5, 10, 25, 50, 100, 250 µg/mL
Experiment I (without activation): 0.05, 0.1, 0.25, 0.5, 1, 2.5, 5, 10, 25, 50, 100 and 200 µg/mL
Experiment I (with activation): 0.5, 1, 2.5, 5, 10, 25, 50, and 100 µg/mL
Experiment I repeat assay (without activation): 10, 25, 50, 60, 70, 80, 90 and 100 µg/mL
The selection of the concentrations was based on data from the pre-experiments. - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: Due to the nature of the test item, it was not possible to prepare a solution of the test item with an appropriate solvent and with the recommended concentration of 2 mg/mL. By lowering the highest concentration to 0.25 mg/mL and based on the results of the solubility test, the best suited vehicle was DMSO (1% v/v).
Controls
- Untreated negative controls:
- yes
- Remarks:
- treatment medium
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- Seeding of the Cultures: Prior to use, cultures were cleansed of pre-existing cells. Two or three day-old exponentially growing stock cultures (more than 50% confluent) were trypsinised at 37°C for 5 minutes. Then the enzymatic digestion was stopped by adding complete culture medium and a single cell suspension was prepared. The trypsine concentration for all subculturing steps was 0.05%. Approximately 10-20e6 cells per concentration, solvent/negative and positive control, were seeded in complete culture medium (MEM supplemented with 10% FBS) in a culture flask, respectively.
Treatment: Approximately 24 hours after seeding the cells were exposed to designated concentrations of the test item either in the presence or absence of metabolic activation in the mutation experiment. After 4 hours exposure the cultures (single cell culture for experiment I and double cell culture for the repetition of experiment I without metabolic activation) were checked for precipitation and the treatment medium containing the test item was removed. The cells were washed twice with PBS, trypsinised and counted with a cell counter. During the following expression period most of the cells were subcultured in complete culture medium (MEM supplemented with 10% FBS) in a sufficient number of cells (at least 2e6 cells per treatment group). In addition, for determination of the relative survival (RS) two 25 cm2 flasks were seeded with approx. 200 cells in complete culture medium for each treatment group. After incubation for an appropriate time (6 - 7 days) colonies were fixed with methanol, stained with Giemsa and counted. Cytotoxicity (relative survival) was calculated based on the cloning efficiency of cells plated immediately after treatment adjusted by any loss of cells during treatment.
Selection: At the end of the expression period (i.e. after 7 to 9 days) for selection the mutants, about 4e5 cells for each treatment group were seeded in cell culture petri dishes (diameter 90 mm) with selective medium containing 11 µg/mL 6-thioguanine (TG) for further incubation (additional 9 - 11 days). The cloning efficiencies (CE) were determined in parallel to the selection of mutants. For each treatment group two 25 cm2 flasks were seeded with approximately 200 cells in complete culture medium to determine the cloning efficiencies after additional 6 - 8 days. After incubation for an appropriate time (9 - 11 days for mutant frequency and 6 - 8 days for CE) colonies were fixed with methanol, stained with Giemsa and counted. The mutant frequency was calculated based on the number of mutant colonies corrected by the cloning efficiency at the time of mutant selection.
Assessment of Cytotoxicity and Mutant Frequency: Cytotoxicity was evaluated by relative survival (RS). The cloning efficiency (CE) of cells plated immediately after treatment was adjusted by any loss of cells during treatment as compared with adjusted cloning efficiency in negative / solvent controls (assigned a survival of 100%). The mutant frequency (MF) is the cloning efficiency of mutant colonies in selective medium divided by the cloning efficiency in non-selective medium measured for the same culture at the time of selection - Evaluation criteria:
- A test chemical is considered to be clearly negative if, in all experimental conditions examined
- none of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control,
- there is no concentration-related increase when evaluated with an appropriate trend-test
- all results are inside the distribution of the historical negative control data
A test chemical is considered to be clearly positive if, in any of the experimental conditions examined
- at least one of the test concentrations exhibits a statistically significant increase compared with the concurrent negative control, and
- the increase is concentration-related when evaluated with an appropriate trend test, and
- any of the results are outside the distribution of the historical negative control data.
- if there is by chance a low spontaneous mutation rate in the corresponding negative and solvent controls a concentration related increase of the mutations within their range has to be discussed.
According to the OECD guideline, the biological relevance is considered first for the interpretation of results. - Statistics:
- The non-parametric Mann-Whitney test was applied to the mutation data to prove the dose groups for any significant difference in mutant frequency compared to the negative/solvent controls. Mutant frequencies of the negative/solvent controls were used as reference.
Results and discussion
Test results
- Key result
- Species / strain:
- Chinese hamster lung fibroblasts (V79)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Exp. I (with and without activation): no growth inhibition observed. Exp. I repeat (without activation): relative survival <70%; relative survival for all concentrations except highest was within range 50-63% (slightly below non-toxic section)
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- Precipitation of the test item was noted in the pre- and main experiment at concentration 100 µg/mL. In the repetition of experiment I precipitation occurred at concentration 80 µg/mL.
For experiment I without metabolic activation the mutant frequencies for the two highest concentrations (50 and 100 µg/mL) were found above the historical data of the test facility. As there was no statistically significant increase, these results needed to be clarified in an independent repetition experiment. In the repetition of experiment I without metabolic activation none of the observed mutant frequencies was significantly increased over those of the negative controls. No dose-response relationship was observed.
DMBA and EMS were used as positive controls and showed distinct and biologically relevant effects in mutation frequency.
Any other information on results incl. tables
Experiment I – Toxicity, without metabolic activation |
|||||||||
Dose Group |
Concentration [µg/mL] |
Number of cells at |
|
Number of colonies per flask |
CE [%] |
Adjusted CE [%] |
Relative Survival (RS) [%] |
||
beginning of treatment |
endo of treatment |
I |
II |
mean |
|
||||
NC1 |
0 |
10000000 |
12665000 |
140 |
165 |
153 |
76 |
97 |
120 |
NC2 |
10000000 |
12818000 |
159 |
172 |
166 |
83 |
106 |
132 |
|
S1 |
0 |
10000000 |
10642000 |
141 |
120 |
131 |
65 |
69 |
100 |
S2 |
10000000 |
10846000 |
166 |
170 |
168 |
84 |
91 |
||
1 |
0.05 |
10000000 |
10625000 |
147 |
150 |
149 |
74 |
79 |
98 |
2 |
0.1 |
10000000 |
9537000 |
157 |
176 |
167 |
83 |
79 |
99 |
3 |
0.25 |
10000000 |
10013000 |
174 |
177 |
176 |
88 |
88 |
109 |
4 |
0.5 |
10000000 |
9622000 |
144 |
158 |
151 |
76 |
73 |
90 |
5 |
1 |
10000000 |
10217000 |
141 |
147 |
144 |
72 |
74 |
92 |
6 |
2.5 |
10000000 |
10591000 |
146 |
152 |
149 |
75 |
79 |
98 |
7 |
5 |
10000000 |
9418000 |
177 |
195 |
186 |
93 |
88 |
109 |
8 |
10 |
10000000 |
9265000 |
178 |
218 |
198 |
99 |
92 |
114 |
9 |
25 |
10000000 |
8993000 |
166 |
170 |
168 |
84 |
76 |
94 |
10 |
50 |
10000000 |
8041000 |
159 |
164 |
162 |
81 |
65 |
81 |
11 P |
100 |
10000000 |
8041000 |
191 |
180 |
186 |
93 |
75 |
93 |
12 P |
200 |
10000000 |
7854000 |
202 |
188 |
195 |
98 |
77 |
95 |
EMS |
300 |
10000000 |
13107000 |
181 |
168 |
175 |
87 |
114 |
142 |
NC: negative control S: solvent control (1% DMSO) P: precipitation at the end of treatment CE: cloning efficiency EMS: Ethylmethanesulfonate |
Experiment I – Mutagenicity, without metabolic activation |
||||||||||||||
|
CE in non-selective medium |
CE in selective medium |
|
|||||||||||
Dose Group |
Concen-tration [µg/mL] |
Number of colonies per flask |
CE [%] |
Number of colonies per flask |
CE [%] |
Mutant Frequency per 106cells |
||||||||
I |
II |
mean |
I |
II |
III |
IV |
V |
mean |
SD |
|||||
NC1 |
0 |
185 |
183 |
184 |
92 |
18 |
11 |
17 |
13 |
14 |
14.6 |
2.6 |
0.0037 |
39.7 |
NC2 |
184 |
186 |
185 |
93 |
7 |
14 |
15 |
9 |
13 |
11.6 |
3.1 |
0.0029 |
31.4 |
|
S1 |
0 |
148 |
187 |
168 |
84 |
14 |
12 |
7 |
17 |
7 |
11.4 |
3.9 |
0.0029 |
34.0 |
S2 |
143 |
186 |
165 |
82 |
19 |
9 |
11 |
13 |
13 |
13.0 |
3.3 |
0.0033 |
39.5 |
|
7 |
5 |
182 |
164 |
173 |
87 |
16 |
13 |
14 |
15 |
11 |
13.8 |
1.7 |
0.0035 |
39.9 |
8 |
10 |
187 |
181 |
184 |
92 |
10 |
14 |
16 |
17 |
10 |
13.4 |
2.9 |
0.0034 |
36.4 |
9 |
25 |
185 |
149 |
167 |
84 |
11 |
15 |
7 |
12 |
16 |
12.2 |
3.2 |
0.0031 |
36.5 |
10 |
50 |
189 |
186 |
188 |
94 |
15 |
15 |
19 |
18 |
21 |
17.6 |
2.3 |
0.0044 |
46.9 |
11 P |
100 |
183 |
160 |
172 |
86 |
12 |
8 |
14 |
26 |
11 |
14.2 |
6.2 |
0.0036 |
41.4 |
EMS |
300 |
134 |
115 |
125 |
62 |
78 |
89 |
92 |
90 |
94 |
88.6 |
5.6 |
0.0222 |
355.8 |
NC: negative control S: solvent control (1% DMSO) P: precipitation at the end of treatment CE: cloning efficiency EMS: Ethylmethanesulfonate |
Experiment I – Toxicity, with metabolic activation |
|||||||||
Dose Group |
Concen-tration |
Number of cells at the |
Number of colonies per flask |
CE [%] |
Adjusted CE [%] |
Relative Survival (RS) |
|||
[µg/mL] |
beginning of treatment |
end of treatment |
I |
II |
mean |
[%] |
|||
NC1 |
0 |
10000000 |
11288000 |
175 |
148 |
162 |
81 |
91 |
97 |
NC2 |
10000000 |
10863000 |
157 |
158 |
158 |
79 |
86 |
91 |
|
S1 |
0 |
10000000 |
11458000 |
143 |
179 |
161 |
81 |
92 |
100 |
S2 |
10000000 |
12257000 |
147 |
168 |
158 |
79 |
97 |
||
1 |
0.5 |
10000000 |
11237000 |
173 |
175 |
174 |
87 |
98 |
104 |
2 |
1 |
10000000 |
11424000 |
154 |
156 |
155 |
78 |
89 |
94 |
3 |
2.5 |
10000000 |
11900000 |
153 |
167 |
160 |
80 |
95 |
101 |
4 |
5 |
10000000 |
11730000 |
125 |
138 |
132 |
66 |
77 |
82 |
5 |
10 |
10000000 |
11305000 |
200 |
203 |
202 |
101 |
114 |
121 |
6 |
25 |
10000000 |
10744000 |
189 |
195 |
192 |
96 |
103 |
109 |
7 |
50 |
10000000 |
10591000 |
204 |
207 |
206 |
103 |
109 |
115 |
8:00 PM |
100 |
10000000 |
9979000 |
177 |
160 |
169 |
84 |
84 |
89 |
DMBA |
1 |
10000000 |
11441000 |
151 |
132 |
142 |
71 |
81 |
86 |
NC: negative control |
Experiment I – Mutagenicity, with metabolic activation |
||||||||||||||
|
CE in non-selective medium |
CE in selective medium |
|
|||||||||||
Dose Group |
Concen-tration [µg/mL] |
Number of colonies per flask |
CE [%] |
Number of colonies per flask |
CE [%] |
Mutant Frequency per 106cells |
||||||||
I |
II |
mean |
I |
II |
III |
IV |
V |
mean |
SD |
|||||
NC1 |
0 |
199 |
215 |
207 |
104 |
21 |
17 |
21 |
19 |
11 |
17.8 |
3.7 |
0.0045 |
43.0 |
NC2 |
166 |
179 |
173 |
86 |
12 |
16 |
14 |
14 |
10 |
13.2 |
2.0 |
0.0033 |
38.3 |
|
S1 |
0 |
174 |
197 |
186 |
93 |
5 |
16 |
8 |
5 |
7 |
8.2 |
4.1 |
0.0021 |
22.1 |
S2 |
189 |
194 |
192 |
96 |
16 |
15 |
15 |
12 |
17 |
15.0 |
1.7 |
0.0038 |
39.2 |
|
4 |
5 |
157 |
173 |
165 |
83 |
15 |
15 |
12 |
14 |
12 |
13.6 |
1.4 |
0.0034 |
41.2 |
5 |
10 |
174 |
167 |
171 |
85 |
9 |
9 |
10 |
10 |
15 |
10.6 |
2.2 |
0.0027 |
31.1 |
6 |
25 |
157 |
154 |
156 |
78 |
10 |
13 |
15 |
10 |
12 |
12.0 |
1.9 |
0.0030 |
38.6 |
7 |
50 |
168 |
162 |
165 |
83 |
8 |
15 |
12 |
11 |
7 |
10.6 |
2.9 |
0.0027 |
32.1 |
8 P |
100 |
157 |
173 |
165 |
83 |
8 |
14 |
8 |
8 |
10 |
9.6 |
2.3 |
0.0024 |
29.1 |
DMBA |
1.0 |
134 |
142 |
138 |
69 |
93 |
101 |
104 |
78 |
81 |
91.4 |
10.4 |
0.0229 |
331.2 |
NC: negative control S: solvent control (1% DMSO) P: precipitation at the end of treatment CE: cloning efficiency DMBA: 7,12-dimethylbenz(a)anthracene |
Experiment I (repetition) – Toxicity, without metabolic activation |
|||||||||
Dose Group |
Concen-tration [µg/mL] |
Number of cells at the |
Number of colonies per flask |
CE [%] |
Adjusted CE [%] |
Relative Survival (RS) [%] |
|||
beginning of treatment |
end of treatment |
I |
II |
mean |
|||||
NC1 |
0 |
10000000 |
9010000 |
73 |
73 |
73 |
37 |
33 |
78 |
NC2 |
10000000 |
10081000 |
92 |
93 |
93 |
46 |
47 |
111 |
|
S1 |
0 |
10000000 |
7633000 |
125 |
128 |
127 |
63 |
48 |
100 |
S2 |
10000000 |
8058000 |
86 |
91 |
89 |
44 |
36 |
||
1 |
10 |
20000000 |
13090000 |
67 |
65 |
66 |
33 |
22 |
51 |
2 |
25 |
20000000 |
11424000 |
81 |
68 |
75 |
37 |
21 |
51 |
3 |
50 |
20000000 |
10914000 |
100 |
84 |
92 |
46 |
25 |
60 |
4 |
60 |
20000000 |
11322000 |
69 |
80 |
75 |
37 |
21 |
50 |
5 |
70 |
20000000 |
11356000 |
90 |
97 |
94 |
47 |
27 |
63 |
6 P |
80 |
20000000 |
12512000 |
104 |
126 |
115 |
58 |
36 |
86 |
7 P |
90 |
20000000 |
13430000 |
142 |
131 |
137 |
68 |
46 |
109 |
8 P |
100 |
20000000 |
12580000 |
120 |
141 |
131 |
65 |
41 |
98 |
EMS |
300 |
10000000 |
11475000 |
139 |
134 |
137 |
68 |
78 |
187 |
NC: negative control S: solvent control (1% DMSO) P: precipitation at the end of treatment CE: cloning efficiency EMS: Ethylmethanesulfonate |
Experiment I (repetition) – Mutagenicity, without metabolic activation |
||||||||||||||
CE in non-selective medium |
CE in selective medium |
|||||||||||||
Dose Group |
Concen-tration [µg/mL] |
Number of colonies per flask |
CE [%] |
Number of colonies per flask |
CE [%] |
Mutant Frequency per 106cells |
||||||||
I |
II |
mean |
I |
II |
III |
IV |
V |
mean |
SD |
|||||
NC1 |
0 |
155 |
160 |
158 |
79 |
8 |
6 |
6 |
11 |
3 |
6.8 |
2.6 |
0.0017 |
21.6 |
NC2 |
147 |
144 |
146 |
73 |
13 |
15 |
13 |
12 |
10 |
12.6 |
1.6 |
0.0032 |
43.3 |
|
S1 |
0 |
133 |
130 |
132 |
66 |
7 |
5 |
8 |
9 |
8 |
7.4 |
1.4 |
0.0019 |
28.1 |
S2 |
152 |
151 |
152 |
76 |
7 |
11 |
11 |
14 |
9 |
10.4 |
2.3 |
0.0026 |
34.3 |
|
2 |
25 |
162 |
158 |
160 |
80 |
14 |
8 |
11 |
8 |
6 |
9.4 |
2.8 |
0.0024 |
29.4 |
3 |
50 |
179 |
165 |
172 |
86 |
6 |
12 |
12 |
5 |
12 |
9.4 |
3.2 |
0.0024 |
27.3 |
4 |
60 |
141 |
152 |
147 |
73 |
7 |
8 |
11 |
15 |
7 |
9.6 |
3.1 |
0.0024 |
32.8 |
5 |
70 |
150 |
140 |
145 |
73 |
12 |
12 |
15 |
5 |
6 |
10.0 |
3.8 |
0.0025 |
34.5 |
6 P |
80 |
169 |
151 |
160 |
80 |
6 |
7 |
7 |
3 |
6 |
5.8 |
1.5 |
0.0015 |
18.1 |
EMS |
300 |
138 |
152 |
145 |
73 |
91 |
80 |
96 |
80 |
86 |
86.6 |
6.2 |
0.0217 |
298.6 |
NC: negative control S: solvent control (1% DMSO) P: precipitation at the end of treatment CE: cloning efficiency EMS: Ethylmethanesulfonate |
Applicant's summary and conclusion
- Conclusions:
- Under the experimental conditions reported, the test item is non-mutagenic at the HPRT locus using V79 cells of the Chinese Hamster.
- Executive summary:
The test substance was assessed for its potential to induce mutations at the HPRT locus using V79 cells of the Chinese Hamster in accordance with OECD Guideline 476. The selection of the concentrations was based on data from the pre-experiments. Experiment I (with and without metabolic activation) and repetition experiment I (without metabolic activation) were performed as a 4 h short-term exposure assay. In consultation with the Sponsor and due to the equivocal results in experiment I, an independent repetition of experiment I without metabolic activation with different experimental conditions (closer spaced concentrations and double cultures) was conducted. The test item was investigated at the following concentrations: 5.0, 10, 25, 50 and 100 µg/mL (Experiment I, without and with metabolic activation) and 25, 50, 60, 70 and 80 µg/mL (Experiment I (repetition), without metabolic activation).
Precipitation of the test item was noted in the pre- and main experiment at concentration 100 µg/mL. In the repetition of experiment I precipitation occurred at concentration 80 µg/mL. No growth inhibition was observed in the experiment I without and with metabolic activation. A growth inhibition (relative survival < 70%) was observed in the repetition experiment without metabolic activation: the relative survival for all concentrations evaluated, with the exception of the highest concentration, was found slightly below the non-toxic section (within the range of 50-63 %). For experiment I without metabolic activation the mutant frequencies for the two highest concentrations (50 and 100 µg/mL) were found above the historical data of the test facility. As there was no statistically significant increase, these results needed to be clarified in an independent repetition experiment. In the repetition of experiment I without metabolic activation none of the observed mutant frequencies was significantly increased over those of the negative controls. No dose-response relationship was observed. DMBA and EMS were used as positive controls and showed distinct and biologically relevant effects in mutation frequency. In conclusion, in the described mutagenicity test under the experimental conditions reported, the test substance is considered to be non-mutagenic at the HPRT locus using V79 cells of the Chinese Hamster.
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.