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EC number: 239-032-7 | CAS number: 14960-06-6
- 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
![](https://echa.europa.eu/o/diss-blank-theme/images/factsheets/A-REACH/factsheet/print_toxicological-information.png)
Endpoint summary
Administrative data
Key value for chemical safety assessment
Genetic toxicity in vitro
Description of key information
Based on the available information, the test item did not cause genotoxic effects in mammalian and bacterial cells under the experimental conditions selected, therefore it is considered not to be mutagenic.
Link to relevant study records
- 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
- Qualifier:
- according to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Deviations:
- no
- GLP compliance:
- yes (incl. QA statement)
- Type of assay:
- mammalian cell gene mutation assay
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): DERIPHAT 160 C
- Physical state:Solid (lyophilized), white
- Analytical purity: 97% The test substance was characterized analytically (for details see the analytical report; project no.: AU 122958-1)
- Lot/batch No.:4986V1
- Expiration date of the lot/batch: until 03 Jun 2014
Homogeneity: The homogeneity of the test substance was ensured by mixing before preparation of the test substance solutions.
- Storage condition of test material:Room temperature (avoid temperatures > 40°C) - Target gene:
- X-linked hypoxanthine-guanine phosphoribosyl transferase (HGPRT) locus
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 mix from phenobarbital- and ß-naphthoflavone induced rats
- Test concentrations with justification for top dose:
- 1st Experiment, without S9 mix: 0; 12.5; 25.0; 50.0; 100.0; 200.0; 400.0 µg/mL
1st Experiment, with S9 mix: 0; 18.8; 37.5; 75.0; 150.0; 300.0; 600.0 µg/mL
2nd Experiment, without S9 mix: 0; 18.8; 37.5; 75.0; 150.0; 300.0; 600.0 µg/mL
2nd Experiment, with S9 mix: 0; 25.0; 50.0; 100.0; 200.0; 400.0; 600.0 µg/mL - Vehicle / solvent:
- Due to the good solubility of the test substance in water, culture medium (Ham's F12) was used as most suitable vehicle.
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- Remarks:
- Negative controls, with and without S9 mix, were treated with culture medium without test substance in parallel to the other treatment groups.
- Positive controls:
- yes
- Positive control substance:
- 7,12-dimethylbenzanthracene
- ethylmethanesulphonate
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Attachment period:20 - 24 hours
- Exposure duration:4 hour, removal of test substance by intense washing (4 hour exposure)
- Expression time (cells in growth medium):3 days (4-hour treatment)
- Selection time (if incubation with a selection agent):6 - 7 days
- Fixation time (start of exposure up to fixation or harvest of cells):10-11 days
SELECTION AGENT (mutation assays): Hypoxanthine-free Ham's F12 medium with 6-thioguanine (10 μg/mL), stable glutamine (200 mM), fetal calf serum (FCS)
STAIN (for cytogenetic assays):colonies were fixed with methanol, stained with Giemsa
NUMBER OF REPLICATIONS:Duplicate cultures were used for all experimental
groups.
NUMBER OF CELLS EVALUATED: Cloning efficiency 200 cells per dose group were seeded in 25 cm² flasks in duplicate using 5 mL. For selection of the mutants, six 75 cm2 flasks with 3x105 cells each from every treatment group, if possible, were seeded in 10 mL selection medium.
DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency - Evaluation criteria:
- The cloning efficiency (CE, %) was calculated for each test group as follows:
CEabsolute = total number of colonies in the test group/total number of seeded cells in the test group x 100
The uncorrected mutant frequency (MFuncorr.) per 106 cells was calculated for each test group
as follows:
MFuncorr. = total number of mutant colonies / number of seeded cells x 106
MFcorr. = MFuncorr. / CE2 absolut x 100
A finding is assessed as positive if the following criteria are met:
• Increase in the corrected mutation frequencies (MFcorr.) both above the concurrent negative control values and our historical negative control data range (see Appendix 6).
• Evidence of the reproducibility of any increase in mutant frequencies.
• A statistically significant increase in mutant frequencies and the evidence of a doseresponse relationship.
The test substance is considered non-mutagenic according to the following criteria:
• The corrected mutation frequency (MFcorr.) in the dose groups is not statistically significantly increased above the concurrent negative control and is within our historical negative control data range. - Statistics:
- An appropriate statistical trend test was performed to assess a dose-related increase of mutant frequencies. The number of mutant colonies obtained for the test substance treated groups was compared with that of the respective negative control groups. A trend is judged as statistically significant whenever the p-value (probability value) is below 0.10 and the slope is greater than 0. However, both, biological and statistical significance will be considered together.
- Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- with S9 in highest tested concentration
- Vehicle controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- In both experiments, in the presence and absence of S9 mix, after 4 hours treatment the morphology and attachment of the cells were adversely influenced in at least the highest applied concentration
- Endpoint:
- in vitro gene mutation study in bacteria
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 29 Oct 2007 - 26 Nov 2007
- 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
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): C-SAT 070070
- Physical state: liquid, light yellow
- Analytical purity: 30% (a.i.)
- Lot/batch No.: S761700005
- Storage condition of test material: room temperature - Target gene:
- his operon for S. typhimurium strains
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98, TA 100 and TA 102
- Additional strain / cell type characteristics:
- other: TA 98: rfa-, uvrB-, R-factor; TA 100: rfa-, uvrB-, R-factor; TA 1535: rfa-, uvrB-; TA 1537: rfa-, uvrB-; TA 102: rfa-; uvrB+. R-factor
- Metabolic activation:
- with and without
- Metabolic activation system:
- S9 liver mix prepared from the liver fraction of Wistar rats treated with Phenobarbital/ß-Naphthoflavone
- Test concentrations with justification for top dose:
- First experiment (initially pre-experiment, with and without metabolic activation): 3, 10, 33, 100, 333, 1000, 2500 and 5000 µg/plate
Second experiment (without metabolic activation): 1, 3, 10, 33, 100, 333, 1000 and 2500 µg/plate
Second experiment (with metabolic activation): 3, 10, 33, 100, 333, 1000, 2500 and 5000 µg/plate - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water
- Untreated negative controls:
- yes
- Remarks:
- (sterility control)
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- with S9-mix
- Positive control substance:
- other: 2-aminoanthracene (2-AA)
- Untreated negative controls:
- yes
- Remarks:
- (sterility control)
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- without S9-mix
- Positive control substance:
- methylmethanesulfonate
- Untreated negative controls:
- yes
- Remarks:
- (sterility control)
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- without S9-mix
- Positive control substance:
- sodium azide
- Untreated negative controls:
- yes
- Remarks:
- (sterility control)
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Remarks:
- without S9-mix
- Positive control substance:
- other: 4-nitro-o-phenylendiamine (NOPD)
- Details on test system and experimental conditions:
- EXPERIMENT I (PRETEST), PLATE INCORPORATION ASSAY
In the standard plate test, tubes were filled with 2 mL portions of overlay agar, 0.1 mL test solution or vehicle or positive control substance, 0.1 mL fresh bacterial culture and 0.5 mL S9 -mix or phosphate buffer. After mixing samples were poured onto selective agar plates and incubated for at least 48 hours in the dark at 37°C.
Triplicate testing is done.
EXPERIMENT II, PREINCUBATION ASSAY
For the preincubation test 0.1 mL test solution or vehicle, 0.1 mL bacterial suspension and 0.5 mL of the S9 mix were mixed and incubated at 37°C for 60 minutes. After addition of 2 mL overlay agar (45°C) samples were poured onto minimal agar plates and incubated for at least 48 hours in the dark at 37°C.
Triplicate testing is done.
DETERMINATION OF CYTOTOXICITY
1.) decrease in the number of revertants
2.) titer reduction
3.) clearing or diminution of the background lawn
TEST CONFOUNDING EFFECTS
As long as no interference between precipitation and colony counting occurs is 5 mg/plate set as maximum dose even for relatively insoluble compounds. - Evaluation criteria:
- An assay is accepted when the following criteria are met:
1.) number of colonies in the negative control is in the historical control range
2.) no indication of bacterial contamination (checked by sterility control)
3.) number of colonies in the positive controls are in the range of historical control data
4.) titer of viable bacteria is ≥ 10 E+9/mL
A test chemical is to be considered as mutagenic when:
An increase of number of revertant colonies (by a factor of 2 for TA 98, TA 100 and TA 102; by a factor of 3 for TA 1535 and TA 1537) is seen, is reproducible and dose-related.
A test chemical is to be considered as non-mutagenic when:
The number of revertants is inside the range of historical negative control data in 2 experiments performed independently from each other. - Key result
- Species / strain:
- S. typhimurium, other: TA 1535, TA 1537, TA 98, TA 100, TA 102
- Remarks:
- Experiment I and Experiment II
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Remarks:
- in none of the experiments, with and without S9 mix, an increase in number of revertants was observed
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- reduced background growth and reduction in number of revertants as cytotoxicity indicators were observed at different concentration levels, in absence and presence of S9 mix.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test item at any dose Level, neither in the presence or absence of metabolic activation (S9 mix).
No tendency of higher mutation rates with increasing concentration in the range below the generally acknowledged border of biological relevance - Endpoint:
- in vitro cytogenicity / chromosome aberration study in mammalian cells
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 20 Dec 1989 - 06 Aug 1990
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- study well documented, meets generally accepted scientific principles, acceptable for assessment
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
- Deviations:
- yes
- Remarks:
- lack of study details, purity of test substance not specified
- GLP compliance:
- not specified
- Type of assay:
- other: in vitro mammalian chromosome aberration test
- Specific details on test material used for the study:
- - Name of test material (as cited in study report): Sodium Laurimindiproprionate (Deriphat 160C)
- Test article ID: (TSIN) MV#2716-040
- Physical state: light yellow liquid
- Analytical purity: not specified
- Storage condition of test material: room temperature protected from light
- Expiration date of the lot/batch: 1990-09-30 - Target gene:
- not applicable
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Metabolic activation system:
- cofactor supplemented post-mitochondrial fraction (S9 mix), prepared from the livers of rats treated with Aroclor 1254
- Test concentrations with justification for top dose:
- with S9:
8 hours harvest - 0.21, 0.28, 2.38 µL/mL
12 hour harvest - 0.28, 0.38, 0.50 µL/mL
without S9:
8 hour harvest: 0.48, 0.63, 0.84 µL/mL
12 hour harvest: 0.63, 0.84, 1.13 µL/mL - Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: water
- Untreated negative controls:
- yes
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- cyclophosphamide
- other: Triethylenemelamine (TEM)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 8 and 12 hours
NUMBER OF REPLICATIONS: 2
NUMBER OF CELLS EVALUATED: 100 per culture
DETERMINATION OF CYTOTOXICITY
- Method: mitotic index - Statistics:
- The cytotoxic effects of treatment are expressed relative to the solvent control (relative cloning efficiency). The number and types of aberrations found are presented for each treatment flask. Duplicate treatment flasks were compared using the Fisher's exact test and, if not statistically significant, combined for treatment comparison.
The percentage of damaged cells (numerical and structural) in the total population of cells examined was calculated for each group. A statistical analysis of the percent aberrant cells per dose was made using the Fisher's exact test.
The average number of aberrations per cell was reported but no statistical analysis was applied. The Cochran-Armitage trend test was performed between the solvent and treatment groups for each treatment condition and harvest time to test for evidence of a dose response. - Key result
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- Relative cloning efficiency at the highest dose level tested of 34% and 1% in the 8 hour and 12 hour non-activated studies, respectively, and 70% and 38% in the 8 hour and 12 hour S9 activated studies, respectively.
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Additional information on results:
- TEST-SPECIFIC CONFOUNDING FACTORS
- Effects of pH: stock concentrations were adjusted to pH7 in order to ensure neutrality of the treatment medium
- Effects of osmolality: osmolality was measured before the main experiments and concentrations chosen where no difference in osmolality was observed
RANGE-FINDING/SCREENING STUDIES:
Dose levels were chosen according to a range-finding toxicity study which was based on cloning efficiency relative to the solvent control. Based upon the findings of the toxicity study dose levels 0.5 and 1.5 µl/mL were the high doses selected for further study in the non-activated and S9 activated studies, respectively.
The average cell generation time (AGT) was calculated in the non-activated and S9 activated studies for the solvent and the two highest test concentrations yielding metaphase cells. At the absence of cell cycle delay harvest times were set at 8 and 12 hours after initiation of treatment for non-activated and S9 activated studies.
Referenceopen allclose all
EXPERIMENT I, with S9 mix: mean number of revertant colonies per dose and strain [3 replicates] |
||||||
Concentration (µg/plate) |
Salmonella typhimurium strain |
|||||
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
TA 102 |
||
untreated |
13 |
20 |
36 |
159 |
495 |
|
Vehicle (water) control |
16 |
21 |
37 |
151 |
575 |
|
TS
|
3 |
18 |
18 |
40 |
145 |
531 |
10 |
18 |
21 |
41 |
152 |
549 |
|
33 |
19 |
24 |
43 |
171 |
568 |
|
100 |
18 |
23 |
38 |
157 |
587 |
|
333 |
20 |
29 |
38 |
165 |
514 |
|
1000 |
15 |
29 |
48 |
148 |
477 |
|
2500 |
12(RB) |
17(RB) |
18(RB) |
90(RB) |
230(RB) |
|
5000 |
1(RB, M) |
1(RB, M) |
0(RB, M) |
15(RB, M) |
0(RB, M) |
|
Positive control (2-AA) |
282 |
322 |
1645 |
2745 |
1842 |
|
EXPERIMENT I, without S9 mix: mean number of revertant colonies per dose and strain [3 replicates] |
||||||
Concentration (µg/plate) |
Salmonella typhimurium strain |
|||||
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
TA 102 |
||
untreated |
19 |
18 |
34 |
142 |
447 |
|
Vehicle (water) control |
15 |
14 |
36 |
150 |
432 |
|
TS
|
3 |
16 |
18 |
30 |
130 |
423 |
10 |
14 |
18 |
30 |
141 |
429 |
|
33 |
11 |
14 |
37 |
129 |
468 |
|
100 |
13 |
17 |
34 |
135 |
429 |
|
333 |
9 |
8(RB) |
18 |
81(RB) |
264(RB) |
|
1000 |
6(RB, M) |
5(RB, M) |
7(RB, M) |
56(RB, M) |
52(RB, M) |
|
2500 |
0(RB, M) |
0(RB, M) |
0(RB, M) |
0(RB, M) |
0(RB, M) |
|
5000 |
0(RB, M) |
0(RB, M) |
0(RB, M) |
0(RB, M) |
0(RB, M) |
|
Respective Positive controls |
2030 |
98 |
421 |
2170 |
3651 |
|
EXPERIMENTII, with S9 mix: mean number of revertant colonies per dose and strain [3 replicates] |
||||||
Concentration (µg/plate) |
Salmonella typhimurium strain |
|||||
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
TA 102 |
||
untreated |
24 |
18 |
39 |
187 |
615 |
|
Vehicle (water) control |
20 |
14 |
41 |
176 |
646 |
|
TS
|
3 |
20 |
16 |
41 |
166 |
608 |
10 |
20 |
13 |
44 |
195 |
658 |
|
33 |
20 |
14 |
41 |
182 |
673 |
|
100 |
21 |
21 |
45 |
171 |
636 |
|
333 |
17 |
20 |
34 |
167 |
624 |
|
1000 |
12(RB) |
23(RB) |
30(RB) |
148 |
536 |
|
2500 |
9(RB) |
14(RB, M) |
8(RB, M) |
106(RB) |
334(RB) |
|
5000 |
2(RB, M) |
0(RB, M) |
0(RB, M) |
0(RB, M) |
3(RB, M) |
|
Positive control (2-AA) |
204 |
161 |
649 |
1733 |
2664 |
|
EXPERIMENT II, without S9 mix: mean number of revertant colonies per dose and strain [3 replicates] |
||||||
Concentration (µg/plate) |
Salmonella typhimurium strain |
|||||
TA 1535 |
TA 1537 |
TA 98 |
TA 100 |
TA 102 |
||
untreated |
16 |
14 |
25 |
150 |
481 |
|
Vehicle (water) control |
14 |
12 |
27 |
153 |
501 |
|
TS
|
1 |
21 |
13 |
30 |
140 |
439 |
3 |
17 |
12 |
29 |
147 |
497 |
|
10 |
15 |
10 |
29 |
131 |
486 |
|
33 |
16 |
13 |
32 |
151 |
503 |
|
100 |
19 |
13 |
34 |
125 |
446 |
|
333 |
15 |
10 |
15 |
106 |
380 |
|
1000 |
8(RB, M) |
3(RB, M) |
2(RB, M) |
14(RB, M) |
277(RB) |
|
2500 |
0(RB, M) |
0(RB, M) |
0(RB, M) |
0(RB, M) |
0(RB, M) |
|
Respective Positive controls |
2006 |
132 |
613 |
2110 |
3510 |
|
TS: test substance; RB: reduced background growth; M: manual count |
Table1: summary of cytogenetic assay results
Treatment1 | Harvest time (hour) | Mean mitotic index2 | Cells scored | Cels with structural aberrations (%)3,4 | Structural aberrations per cell (mean ± SD)5 |
Non-activated study | |||||
untreated cells | 8 | 3.4 | 100 | 0 | 0 .000 ± 0.000 |
water | 8 | 4.1 | 100 | 1 | 0.010 ± 0.100 |
0.21 µL/mL | 8 | 3.9 | 100 | 0 | 0.000 ± 0.000 |
0.28 µL/mL | 8 | 2.2 | 100 | 1 | 0.010 ± 0.100 |
0.38 µL/mL | 8 | 0.2 | 67 | 0 | 0.000 ± 0.000 |
untreated cells | 12 | 4.2 | 100 | 0 | 0.000 ± 0.000 |
water | 12 | 3.5 | 100 | 0 | 0.000 ± 0.000 |
0.28 µL/mL | 12 | 2.5 | 100 | 1 | 0.010 ± 0.100 |
0.38 µL/mL | 12 | 0.8 | 100 | 2 | 0.020 ± 0.141 |
0.50 µL/mL | 12 | 0.2 | 64 | 1 | 0.016 ± 0.125 |
TEM, 0,25 µg/mL | 12 | 0.9 | 100 | 11 | 0.130 ± 0.393 |
S9 activated study |
|||||
untreated cells | 8 | 2.5 | 100 | 1 | 0.010 ± 0.100 |
water | 8 | 2.3 | 100 | 3 | 0.030 ± 0.171 |
0.48 µL/mL | 8 | 1.9 | 100 | 1 | 0.010 ± 0.100 |
0.63 µL/mL | 8 | 2.1 | 100 | 1 | 0.010 ± 0.100 |
0.84 µL/mL | 8 | 0.8 | 100 | 1 | 0.010 ± 0.100 |
untreated cells | 12 | 3.7 | 100 | 1 | 0.010 ± 0.100 |
water | 12 | 3.3 | 100 | 1 | 0.010 ± 0.100 |
0.63 µL/mL | 12 | 3.7 | 100 | 3 | 0.030 ± 0.171 |
0.84 µL/mL | 12 | 4.1 | 100 | 2 | 0.020 ± 0.141 |
1.13 µL/mL | 12 | 0.9 | 55 | 0 | 0.000 ± 0.000 |
CP, 20µg/mL | 12 | 1.8 | 100 | 12 | 0.130 ± 0.367 |
1 CHO cells were treated at 37°C for 4 hours
2 Mitotic Index = cells in mitosis per 500 cells counted, expressed as a percentage
3 Structural aberrations include unanalyzable cells and cells with one ore more aberrations, excluding gaps
4 significantly increased above the control using Fisher's exact test; *p<0.025
5 Gaps and unanalyzable cells are not included; 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
In vitro gene mutation study in bacteria:
A study, according to OECD 471 (1997) was performed to investigate the potential of the test item to induce gene mutations according to the plate incorporation test (experiment I) and the pre-incubation test (experiment II) using the Salmonella typhimurium strains TA 1535, TA 1537, TA 98. TA 100, and TA 102 (RCC, 2008). The assay was performed in two independent experiments, with and without liver micmosomal activatiion(S9 mix). Each concentration, including the controls, was tested in triplicate. The test item was tested at the following concentrations of the active ingredient: Pre-Experiment/Experiment I: 3; 10; 33; 100; 333; 1000: 2500: and 5000 µg/plate, Experiment II without S9 mix: 1; 3; 10; 3: 100: 333; 1000; and 2500 µg/plate, Experiment II with S9 mix: 3; 10; 33; 100; 333; 100: 2500, and 5000 µg/plate. The plates incubated with the test item showed reduced background growth with and without S9 mix in all strains used in both experiments. Toxic effect, evident as a reduction in the number of revertents, occurred in the test groups with and without metabolic activation.
No substantial increase in revertant colony numbers of any of the five tester strains was observed following treatment with the test item at any dose Level, neither in the presene or absence of metabolic activation (S9 mix). There was also no tendency of higher mutation rates with increasing concentration in the range below the generally acknowledged border of biological relevance. Appropriate reference mutagens were used as positive controls and showed a distinct increase of induced revertant colonies. Summarizing, the test item is considered not to be mutagenic in this bacterial mutagenicity test in vitro.
In vitro gene mutation study in mammalian cells:
The test substance was assessed for its potential to induce gene mutations at the hypoxanthine-guanine phosphoribosyl transferase (HPRT) locus in Chinese hamster ovary (CHO) cells in vitro according to OECD 476 (BASF SE, 2013). Two independent experiments were carried out, with and without the addition of liver S9 mix from phenobarbital- and β-naphthoflavone induced rats (exogenous metabolic activation). According to an initial range-finding cytotoxicity test for the determination of the experimental doses and taking into account the cytotoxicity actually found in the main experiments, the following doses were tested. 1st and 2nd Experiment without S9 mix 0; 12.5; 25.0; 50.0; 100.0; 200.0; 400.0 μg/mL and with S9 mix 0; 18.8; 37.5; 75.0; 150.0; 300.0; 600.0 μg/mL. Following attachment of the cells for 20-24 hours, cells were treated with the test substance for 4 hours in the absence and presence of metabolic activation. Subsequently, cells were cultured for 6-8 days and then selected in 6-thioguanine-containing medium for another week. Finally, the colonies of each test group were fixed with methanol, stained with Giemsa and counted. The negative controls gave mutant frequencies within the range expected for the CHO cellline. Both positive control substances, EMS and DMBA, led to the expected increase in the frequencies of forward mutations. In this study, in the 1st and 2nd Experiment, the highest concentrations evaluated for gene mutations were clearly cytotoxic in the absence and the presence of metabolic activation. Based on the results of the present study, the test substance did not cause any relevant increase in the mutant frequencies either without S9 mix or after the addition of a metabolizing system in two experiments performed independently of each other. Thus, under the experimental conditions of this study, the test substance is not mutagenic in the HPRT locus assay under in vitro conditions in CHO cells in the absence and the presence of metabolic activation. Summarizing, the test item is not considered to be mutagenic in a gene mutation test in vitro with mammalian cells.
In vitro cytogenicity / chromosome aberration study in mammalian cells:
The test item was tested in the chromosome aberration assay using Chinese hamster ovary cells similar to OECD 473 (CIR, 1990). The assay was conducted both in the absence and presence of an Aroclor induced rat liver S9 activation system. Dose levels of 0.21, 0.28 and 0.38 µl/mL in the 8 hour non-activated study, 0.28, 0.38 and 0.5 µl/mL in the 12 hour non-activated study, 0.48, 0.63 and 0.84 µl/mL in the 8 hour S9 activated study and dose levels of 0.63, 0.84 and 1.13 µl/mL in the 12 hour S9 activated study were selected at the highest, non-precipitating concentrations in the non-activated and S9 activated studies which could be evaluated for chromosome aberrations. Survival (relative cloning efficiency) at the highest dose level scored was 34% and 1% in the 8 and 12 hour non-activated studies, respectively, and 70% and 38% in the 8 and 12 hour S9 activated studies, respectively.Summarizing the test item did not induce a significant increase in chromosome aberrations at either harvest time, 8 or 12 hours, in the absence or presence of S9 activation. Therefore the test substance is considered negative in the CHO cytogenicity study.
Assessment of genetic toxicity:
According to the test strategy for mutagenicity mentioned in Annex VII-IX of REGULATION (EC) No 1907/2006 first in vitro test to investigate mutagenicity has to be performed. Only in cases were evidence from in vitro test (or other information) exist testing in mammals is suggested.
All in vitro tests conducted showed negative results. Therefore the test substance was considered as not mutagenic.
As no indication of mutagenicity can be observed in the available test results and also no other further information gave hints for mutagenicity no additional testing is suggested.
Justification for classification or non-classification
Based on the available information, classification for genetic toxicity is not warranted in accordance with EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation 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.
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