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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.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: 225-806-1 | CAS number: 5089-72-5
- 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
- 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
Genetic toxicity in vitro:
Mutagenicity in bacteria (OECD 471, Ames): negative with and without metabolic activation in Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537.
Cytogenicity in mammalian cells: testing not required as a reliable in vivo micronucleus test is available on the structural analogue substance CAS 1760-24-3.
Mutagenicity in mammalian cells (similar to OECD 476, MLA): negative with and without metabolic activation in CHO cells (RA from CAS 1760-24-3)
Link to relevant study records
- Endpoint:
- in vitro gene mutation study in bacteria
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1994-10-19 to 1994-11-07
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: The study was conducted according to an appropriate OECD test guideline, with acceptable restrictions. The restrictions were [4 strains only. Non-standard positive controls. only 2-aminoanthracene as +S9 control]
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 471 (Bacterial Reverse Mutation Assay)
- Version / remarks:
- 1983
- Deviations:
- yes
- Remarks:
- 4 strains only. Non-standard positive controls. only 2-aminoanthracene as +S9 control
- GLP compliance:
- yes
- Type of assay:
- bacterial reverse mutation assay
- Target gene:
- His operon
- Species / strain / cell type:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor induced rat liver S9
- Test concentrations with justification for top dose:
- 33.3, 100, 333.3, 1000, 2500 and 5000 µg/pate with and without metabolic activation
- Vehicle / solvent:
- - Vehicle(s)/solvent(s) used: ethanol
- Justification for choice of solvent/vehicle: Solubility properties and relative non-toxicity to bacteria - Untreated negative controls:
- yes
- Remarks:
- Untreated culture
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- sodium azide
- Remarks:
- TA 1535, TA 100 (without activation)
- Untreated negative controls:
- yes
- Remarks:
- Untreated culture
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 4-nitro-o-phenylene-diamine
- Remarks:
- TA 1537, TA 98 (without activation)
- Untreated negative controls:
- yes
- Remarks:
- Untreated culture
- Negative solvent / vehicle controls:
- yes
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- other: 2-aminoanthracene
- Remarks:
- All strains (with activation)
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in agar (plate incorporation); preincubation
DURATION
- Preincubation period: 60 minutes
- Exposure duration: 48 hours
NUMBER OF REPLICATIONS: 3 plates for each test concentration
DETERMINATION OF CYTOTOXICITY
- Method: background lawn assessment - Evaluation criteria:
- A test article is considered mutagenic if in the strain TA 100 the number of reversions is at least twice as high and in strains TA 1535, TA 1537 and TA 98 at least three times higher as compared to the spontanious reversion rate. A dose dependant and reproducible increase in the number of revertants is an indication of possible existing mutagenic potential regardless of highest dose value.
- Species / strain:
- S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- - S9 (pre-incubation test): TA 98 and TA1537 at 5000 µg/plate, while TA 100 at 2500 µg/plate. + S9 (pre-incubation test): TA 1537 at 5000 µg/plate
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- negative with metabolic activation
negative without metabolic activation
Under test conditions, no mutagenic effect was observed for the test substance tested up to limit concentration in any of the test strains in two independent experiments without and with metabolic activation. The test substance is non-mutagenic in test strains used. - Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1988-01-13 to 1988-03-21
- Reliability:
- 1 (reliable without restriction)
- Rationale for reliability incl. deficiencies:
- other: GLP guideline study
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
- Qualifier:
- according to guideline
- Guideline:
- other: EPA Health Effects Test Guidelines HG-Gene Muta-Somatic Cells (1983)
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- mammalian cell gene mutation assay
- Target gene:
- HGPRT (hypoxanthine-guanine phosphoribosyl transferase)
- Species / strain / cell type:
- Chinese hamster Ovary (CHO)
- Details on mammalian cell type (if applicable):
- - Type and identity of media: Ham's Modified F12 Medium
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: no
- Periodically "cleansed" against high spontaneous background: no - Additional strain / cell type characteristics:
- not specified
- Metabolic activation:
- with and without
- Metabolic activation system:
- Aroclor-1254 induced rat liver S9
- Test concentrations with justification for top dose:
- without metabolic activation: 2.5, 3.0, 3.25, 3.5 and 4.0 mg/ml, with metabolic activation: 2.0, 2.5, 3.0, 3.5, 4.0 and 4.5 mg/ml
- Vehicle / solvent:
- DMSO
- Untreated negative controls:
- yes
- Remarks:
- cell culture medium
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- ethylmethanesulphonate
- Remarks:
- without metabolic activation
- Untreated negative controls:
- yes
- Remarks:
- cell culture medium
- Negative solvent / vehicle controls:
- yes
- Remarks:
- DMSO
- True negative controls:
- no
- Positive controls:
- yes
- Positive control substance:
- N-dimethylnitrosamine
- Remarks:
- with metabolic activation
- Details on test system and experimental conditions:
- METHOD OF APPLICATION: in medium
DURATION
- Exposure duration: 5 hours (with and without metabolic activation)
- Expression time (cells in growth medium): 9-12 days
- Selection time: 18 and 24 hours
SELECTION AGENT (mutation assays): TG selective medium
NUMBER OF REPLICATIONS: Duplicate cultures. Surviving fraction was determined at 18 and 24 hours after removal of chemical using 4 plates per culture and 100 cells per plate; mutant frequency was determined using 5 plates per culture. The test was repeated.
DETERMINATION OF CYTOTOXICITY
- Method: plating efficiency
ACTIVATION: S9 mix contained glucose-6-phosphate and NADP as co-factors, and 5% of S9. 1.0 ml of S9 mix was added to 4 ml of culture medium, giving a final concentration of 1% S9. - Evaluation criteria:
- The criteria for interpretation of the test results as a positive or negative response depend upon both the level of statistical significance from the concurrent control and the evidence of a dose-response effect following treatment. When a definite dose-response relationship is not evident, but one or more marginally significant values are obtained, a careful examination of the data from the concurrent positive and negative controls and comparisons to historical control data may be used to evaluate the probable biological significance of the responses.
- Statistics:
- Box-Cox transformation
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 4.0 mg/ml with metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Conclusions:
- negative with and without metabolic activation
The test material has been tested in a reliable study according to an appropriate US EPA test guideline that is equivalent to OECD 476 and under GLP. An increase in mutant frequency was observed in one of two duplicate cultures with metabolic activation at one concentration in the initial experiment. No other increase in mutant frequency was observed at any concentration with and without activation (5 hours treatment). The experiment was repeated (with metabolic activation): no increase in mutant frequency was observed. The appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of mutations in CHO cells under the conditions of the test. - Endpoint:
- in vitro gene mutation study in mammalian cells
- Remarks:
- Type of genotoxicity: gene mutation
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Please refer to the Additional Information field in the endpoint study summary.
- Reason / purpose for cross-reference:
- read-across source
- Species / strain:
- Chinese hamster Ovary (CHO)
- Metabolic activation:
- with and without
- Genotoxicity:
- negative
- Cytotoxicity / choice of top concentrations:
- cytotoxicity
- Remarks:
- 4.0 mg/ml with metabolic activation
- Vehicle controls validity:
- valid
- Untreated negative controls validity:
- valid
- Positive controls validity:
- valid
- Remarks on result:
- other: BRRC, 1988
- Conclusions:
- negative with and without metabolic activation
The test material has been tested in a reliable study according to an appropriate US EPA test guideline that is equivalent to OECD 476 and under GLP. An increase in mutant frequency was observed in one of two duplicate cultures with metabolic activation at one concentration in the initial experiment. No other increase in mutant frequency was observed at any concentration with and without activation (5 hours treatment). The experiment was repeated (with metabolic activation): no increase in mutant frequency was observed. The appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of mutations in CHO cells under the conditions of the test.
Referenceopen allclose all
Table 2: Dose range-finding study number of revertants per plate (2 plates per strain)
|
TA 98 |
TA 100 |
||||
Concentration (μg/Plate) |
Plate 1 + MA |
Plate 2 - MA |
Cytotoxic (Yes/No) |
Plate 1 + MA |
Plate 2 - MA |
Cytotoxic (Yes/No) |
Negative control |
15 |
17 |
No |
163 |
122 |
No |
0 |
24 |
16 |
No |
147 |
129 |
No |
3.3 |
12 |
17 |
No |
150 |
128 |
No |
10 |
22 |
20 |
No |
154 |
116 |
No |
33.3 |
18 |
12 |
No |
158 |
120 |
No |
100 |
19 |
19 |
No |
155 |
119 |
No |
333.3 |
21 |
14 |
No |
87 |
107 |
No |
1000 |
23 |
16 |
No |
159 |
109 |
No |
2500 |
20 |
13 |
No |
128 |
111 |
No |
5000 |
57 |
10 |
No |
126 |
111 |
No |
Positive control |
555 |
142 |
No |
973 |
353 |
No |
*solvent control with ethanol
Table 3: Experiment 1 Mutagenicity Assay (plate incorporation) number of revertants per plate (mean of 3 plates)
|
TA98 |
TA100 |
TA1535 |
||||||
Conc. |
— MA |
+ MA |
Cytotoxic |
— MA |
+ MA |
Cytotoxic |
— MA |
+ MA |
Cytotoxic |
Negative control |
17 |
15 |
No |
122 |
163 |
No |
13 |
13 |
No |
0 |
16 |
24 |
No |
129 |
147 |
No |
14 |
19 |
No |
33.3 |
12 |
18 |
No |
120 |
158 |
No |
15 |
16 |
No |
100 |
19 |
19 |
No |
119 |
155 |
No |
15 |
16 |
No |
333.3 |
14 |
21 |
No |
107 |
87 |
No |
13 |
15 |
No |
1000 |
16 |
23 |
No |
109 |
159 |
No |
12 |
20 |
No |
2500 |
13 |
20 |
No |
111 |
128 |
No |
11 |
19 |
No |
5000 |
10 |
57 |
No |
111 |
126 |
No |
12 |
16 |
No |
Positive control |
142 |
555 |
No |
353 |
973 |
No |
69 |
405 |
No |
*solvent control with ethanol
Table 3: Experiment 1 Mutagenicity Assay (plate incorporation ) number of revertants per plate (mean of 3 plates)
|
TA1537 |
||
Conc. |
— MA |
+ MA |
Cytotoxic |
Negative control |
25 |
29 |
No |
0 |
19 |
24 |
No |
33.3 |
17 |
21 |
No |
100 |
18 |
23 |
No |
333.3 |
18 |
21 |
No |
1000 |
20 |
22 |
No |
2500 |
16 |
22 |
No |
5000 |
14 |
22 |
No |
Positive control |
79 |
279 |
No |
*solvent control with ethanol
Table 4: Experiment 2 Mutagenicity Assay (pre-incubation) Number of revertants per plate (mean of 3 plates)
|
TA98 |
TA100 |
TA1535 |
||||||
Conc. |
— MA |
+ MA |
Cytotoxic |
— MA |
+ MA |
Cytotoxic |
— MA |
+ MA |
Cytotoxic |
Negative control |
27 |
39 |
No |
130 |
136 |
No |
10 |
13 |
No |
0 |
31 |
38 |
No |
103 |
122 |
No |
17 |
19 |
No |
33.3 |
27 |
36 |
No |
92 |
129 |
No |
12 |
19 |
No |
100 |
28 |
42 |
No |
105 |
126 |
No |
11 |
15 |
No |
333.3 |
29 |
32 |
No |
94 |
122 |
No |
12 |
19 |
No |
1000 |
22 |
37 |
No |
71 |
142 |
No |
15 |
20 |
No |
2500 |
18 |
31 |
No |
9 |
91 |
Yes |
10 |
13 |
No |
5000 |
0 |
31 |
Yes |
0 |
70 |
Yes |
11 |
8 |
No |
Positive control |
218 |
287 |
No |
808 |
389 |
No |
862 |
122 |
No |
*solvent control with Ethanol
Table 4: Experiment 2 Mutagenicity Assay (pre-incubation) number of revertants per plate (mean of 3 plates)
|
TA1537 |
||
Conc. |
— MA |
+ MA |
Cytotoxic |
Negative control |
29 |
31 |
No |
0 |
29 |
32 |
No |
33.3 |
27 |
36 |
No |
100 |
26 |
35 |
No |
333.3 |
28 |
33 |
No |
1000 |
27 |
35 |
No |
2500 |
19 |
25 |
No |
5000 |
8 |
4 |
Yes |
Positive control |
94 |
121 |
No |
*solvent control with ethanol
MA: Metabolic activation
CHO Mutation Assay: with and without metabolic activation
Results on evaluation of plating efficiency and mutant colonies - Experiment 1
|
Mutant colonies |
Plating efficiency |
||
Test chemical |
(Total of 2 plates) |
% of Combined Solvent Controls |
||
|
- S9 |
+S9 |
-S9 |
+S9 |
Negative control |
0 |
18 |
106.3 |
105.9 |
Solvent control A |
20 |
2 |
93.7 |
102.3 |
Solvent control B |
0 |
0 |
106.3 |
97.8 |
2.0A |
- |
6 |
- |
120.5 |
2.0B |
- |
13 |
- |
104.4 |
2.5A |
6 |
35 |
103.4 |
107.5 |
2.5B |
1 |
15 |
110.0 |
100.5 |
3.0A |
5 |
4 |
104.6 |
91.0 |
3.0B |
2 |
0 |
124.4 |
109.2 |
3.25A |
5 |
- |
101.1 |
- |
3.25B |
5 |
- |
96.0 |
- |
3.5A |
0 |
0 |
89.4 |
116.0 |
3.5B |
1 |
0 |
103.7 |
107.7 |
4.0A |
0 |
-** |
114.7 |
-** |
4.0B |
5 |
-** |
94.0 |
-** |
Positive control |
143 |
55 |
115.7 |
113.2 |
**= Cytotoxic, cultures did not grow
Results on evaluation of plating efficiency and mutant colonies - Experiment 2
Test chemical
|
Mutant colonies (Total of 2 plates) |
Plating efficiency |
+S9 |
||
Negative control |
13 |
88.2 |
Solvent control A |
0 |
104.1 |
Solvent control B |
1 |
95.8 |
2.25A |
12 |
118.7 |
2.25B |
0 |
99.7 |
2.5A |
0 |
106.8 |
2.5B |
0 |
111.1 |
2.75A |
0 |
101.9 |
2.75B |
0 |
96.6 |
Positive control |
63 |
92.6 |
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Genetic toxicity in vivo
Description of key information
Genetic toxicity in vivo:
Micronucleus assay in mouse (EPA 560/6-83-001, similar to OECD 474): negative (RA from CAS 1760-24-3)
Link to relevant study records
- Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Remarks:
- Type of genotoxicity: chromosome aberration
- Type of information:
- experimental study
- Adequacy of study:
- key study
- Study period:
- 1988-03-15 to 1988-04-08
- Reliability:
- 2 (reliable with restrictions)
- Rationale for reliability incl. deficiencies:
- other: see 'Remark'
- Remarks:
- The study was conducted according to a test protocol that is similar to the appropriate OECD test guideline, and in compliance with GLP. According to the ECHA guidance document “Practical guide 6: How to report read-across and categories (March 2010)”, the reliability was changed from RL1 to RL2 to reflect the fact that this study was conducted on a read-across substance (CAS 1760-24-3). Only 1000 cells/animal were scored for micronuclei.
- Qualifier:
- equivalent or similar to guideline
- Guideline:
- OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
- Deviations:
- yes
- Remarks:
- number of micronucleated PCEs scored per 1000 cells, not 2000
- Qualifier:
- according to guideline
- Guideline:
- other: EPA Health Effect T G, EPA Report 560/6-83-001
- Deviations:
- no
- GLP compliance:
- yes
- Type of assay:
- micronucleus assay
- Species:
- mouse
- Strain:
- Swiss Webster
- Sex:
- male/female
- Details on test animals or test system and environmental conditions:
- TEST ANIMALS
- Source: Charles River Laboratories, Portage, MI, USA
- Age at study initiation: 5 weeks
- Weight at study initiation: male - 22.5 g - 28.0 g. female - 19.8 g - 21.7 g
- Assigned to test groups randomly: randomised separately by sex
- Fasting period before study: no
- Housing: 5 mice/sex/cage in shoe-box type plastic cages
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
ENVIRONMENTAL CONDITIONS
- Temperature (°C): controlled but not specified
- Humidity (%): controlled but not specified
- Air changes (per hr): not known
- Photoperiod (hrs dark / hrs light): 12/12 - Route of administration:
- intraperitoneal
- Vehicle:
- - Vehicle(s)/solvent(s) used: corn oil
- Justification for choice of solvent/vehicle: incompatible with water thus corn oil was used
- Concentration of test material in vehicle: 200 mg/kg - Duration of treatment / exposure:
- not applicable
- Frequency of treatment:
- One treatment only
- Post exposure period:
- 30, 48 and 72 hours
- Dose / conc.:
- 87.5 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 175 mg/kg bw/day (actual dose received)
- Dose / conc.:
- 280 mg/kg bw/day (actual dose received)
- No. of animals per sex per dose:
- 5
- Control animals:
- yes
- Positive control(s):
- Triethylenemelamine
- Justification for choice of positive control(s): known to demonstrate the sensitivity and responsiveness of the animals in the definitive test.
- Route of administration: i p
- Doses / concentrations: 0.3 mg/kg bw - Tissues and cell types examined:
- Blood was collected by nicking the tail of each animal with a scalpel and slides prepared for each animal per sampling time. Polychromatic erythrocytes (PCE's) were examined.
- Details of tissue and slide preparation:
- CRITERIA FOR DOSE SELECTION: Determination of the PCE/NCE ratio for the groups of animals with partial mortality was used to evaluate the possibility of bone marrow cytotoxicity from the test chemical. Three dose levels of approximately 80%, 50% and 25% of the LD50 value were evaluated for effects upon the incidence of micronuclei.
DETAILS OF SLIDE PREPARATION: Slides were stained with Gurr's R-66 Giemsa diluted in phosphate buffer. Slides were coded by animal number only and read blindly.
METHOD OF ANALYSIS: A minimum of 1000 PCE's were examined microscopically for each animal per sample time. The PCE/NCE ratio for approximately 1000 total cells was calculated and recorded. The number of micronucleated PCE/1000 NCE was recorded. - Evaluation criteria:
- A test result is considered to be negative if no statistically significant or dose related increases are apparent between the vehicle control and groups of animals treated with Organofunctional Silane A-1120.
- Statistics:
- Fisher's Exact Test (Sokal and Rohlf, 1981)
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- PCE/NCE ratio was reduced at highest concentration, 48 h exposure and at all doses, 72 h exposure.
- Vehicle controls validity:
- valid
- Negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Additional information on results:
- RESULTS OF RANGE-FINDING STUDY
- Dose range: 125 mg/kg bw - 2000 mg/kg b
- Solubility: Incompatible with water thus corn oil was used
- Clinical signs of toxicity in test animals: All mice dosed at 1000 mg/kg bw and 2000 mg/kg bw died. 500 mg/kg bw was lethal to 4 out of 5 females and all the male mice. One female tested at 250 mg/kg bw died.
- Evidence of cytotoxicity in tissue analyzed: a decrease in the PCE/NCE ratio to 80% of the control value was observed at the 48 hour treatment interval. At 72 hours the PCE/NCE ratios had increased.
- Harvest times: 48 hours
RESULTS OF DEFINITIVE STUDY
- Induction of micronuclei (for Micronucleus assay): negative.
- Ratio of PCE/NCE (for Micronucleus assay): PCE/NCE ratio was slightly decreased at 72 hour treatment interval.
- Appropriateness of dose levels and route: Both dose levels and route were appropriate.
- Statistical evaluation: Analysis of variance indicated no sex-related differences in the incidence of micronuclei. No statistically significant (p ≤ 0.01) or treatment related increases in the numbers of micronuclei were observed at any dose or treatment interval. - Conclusions:
- negative
The test material has been tested in a reliable study according to EPA Health Effect T G, Report 560/6-83-001, which is similar to OECD 474, and under GLP conditions. No treatment related increases in numbers of micronuclei in PCE's of Swiss-Webster mice were observed at concentrations of 87.5, 175 and 280 mg/kg bw. Relatively high dose levels of the test compound were tested up to 80% of the LD50 with no indication of significant induction of micronuclei. The PCE/NCE ratio was reduced at the highest concentration, 48 h exposure and at all doses, 72 h exposure, indicating exposure of the target tissue to the test substance. The test compound is considered to be negative for the induction of micronuclei under the conditions of this test. - Endpoint:
- in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
- Remarks:
- chromosome aberration
- Type of information:
- read-across from supporting substance (structural analogue or surrogate)
- Adequacy of study:
- key study
- Justification for type of information:
- Please refer to the Additional Information field in the endpoint study summary.
- Reason / purpose for cross-reference:
- read-across source
- Sex:
- male/female
- Genotoxicity:
- negative
- Toxicity:
- yes
- Remarks:
- PCE/NCE ratio was reduced at highest concentration, 48 h exposure and at all doses, 72 h exposure.
- Vehicle controls validity:
- valid
- Negative controls validity:
- not applicable
- Positive controls validity:
- valid
- Remarks on result:
- other: BRRC, 1988
- Conclusions:
- negative
The test material has been tested in a reliable study according to EPA Health Effect T G, Report 560 /6-83-001, which is similar to OECD 474, and under GLP conditions. No treatment related increases in numbers of micronuclei in PCE's of Swiss-Webster mice were observed at concentrations of 87.5, 175 and 280 mg/kg bw. Relatively high dose levels of the test compound were tested up to 80% of the LD50 with no indication of significant induction of micronuclei. The PCE/NCE ratio was reduced at the highest concentration, 48 h exposure and at all doses, 72 h exposure, indicating exposure of the target tissue to the test substance. The test compound is considered to be negative for the induction of micronuclei under the conditions of this test.
Referenceopen allclose all
Table 3: Summary of micronucleus results
|
30 hours |
48 hours |
72 hours |
|||||||||
Treatment groups mg/kg bw |
Number of PCE’s with micronuclei per 1000 PCE’s |
Mean PCE’s/1000 NCE (SD) |
Number of PCE’s with micronuclei per 1000 PCE’s |
Mean PCE’s/1000 NCE (SD) |
Number of PCE’s with micronuclei per 1000 PCE’s |
Mean PCE’s/1000 NCE (SD) |
||||||
Male |
Female |
Group mean |
SD |
Male |
Female |
Group mean |
SD |
Male |
Female |
Group mean |
SD |
|
Vehicle control |
4.0 (2.00) |
1.6 (0.89) |
33.7 |
5.52 |
2.4 (2.70) |
3.6 (1.14) |
36.4 |
4.24 |
3.8 (2.05) |
3.8 (2.68) |
42.9 |
3.25 |
87.5 |
5.0 (2.55) |
3.6 (1.52) |
31.8 |
6.79 |
3.0 (1.73) |
2.2 (0.84) |
36.0 |
0.28 |
6.4 (2.19) |
2.8 (1.64) |
33.9 |
2.12 |
175 |
2.6 (1.52) |
4.0 (1.22) |
34.9 |
8.06 |
3.2 (1.10) |
3.8 (1.64) |
37.2 |
7.92 |
2.3 (0.96) |
5.2 (3.96) |
35.5 |
6.68 |
280 |
3.4 (2.19) |
3.6 (2.19) |
35.4 |
4.81 |
3.4 (3.29) |
3.0 (1.22) |
30.4 |
3.39 |
3.8 (1.92) |
1.2 (0.84) |
27.9 |
1.27 |
Positive control |
36.2 (16.84) |
28.6 (10.31) |
27.4 |
9.05 |
Not evaluated |
Not evaluated |
Not evaluated |
Not evaluated |
Table 4 Summary of micronucleus frequency and statistical differences from controls (Fisher's exact test, one-tailed)
Treatment/dose mg/kg bw |
No. PCE observed |
PCE with micronuclei |
Statistical significance |
|
30 hour sample |
||
Vehicle control |
10 000 |
28 |
NS |
87.5 |
10 000 |
43 |
0.05>p>0.01* |
175 |
10 000 |
33 |
NS |
280 |
10 000 |
35 |
NS |
Positive control |
10 000 |
324 |
p<0.001 |
|
48 hour sample |
||
Vehicle control |
10 000 |
30 |
NS |
87.5 |
10 000 |
26 |
NS |
175 |
10 000 |
35 |
NS |
280 |
10 000 |
32 |
NS |
|
72 hour sample |
||
Vehicle control |
10 000 |
38 |
NS |
87.5 |
10 000 |
46 |
NS |
175 |
10 000 |
35 |
NS |
280 |
10 000 |
25 |
NS |
Results from sexes are combined as no sex-related differences were shown.
* not considered of biological significance
Endpoint conclusion
- Endpoint conclusion:
- no adverse effect observed (negative)
Additional information
The mutagenicity potential of N-[3-(triethoxysilyl)propyl]ethylenediamine in bacteria was assessed in an experiment according to OECD 471 with Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100 (RCC, 1995). A deviation to the guideline was the use of 2-aminoanthracene as the sole positive control substance in the presence of S9-mix and only 4 strains of Salmonella typhimurium were used. No further information is given, if the S9-mix has been characterised with a second substance. This deviation was supposed to have no implication on the outcome of the study results. The tester strains were treated using both the pre-incubation and the plate incorporation methods, with and without S9-mix, respectively. The concentrations tested were 33.3 - 5000 µg/plate in the presence and absence of S9-mix. Results achieved with vehicle (ethanol) and positive controls were valid. No genotoxicity was observed in the presence and absence of metabolic activation. Growth inhibition was observed in some of the test groups with and without metabolic activation at 2500 and 5000 µg/plate in the pre-incubation test. In conclusion, N-[3-(triethoxysilyl)propyl]ethylenediamine did not induce mutations in bacteria under the test conditions applied.
No data are available for mutagenicity to a bacterial strain capable of detecting cross-linking or oxidising mutagens. In view of the lack of genetic toxicity demonstrated in studies on mammalian cells, and the absence of structural features that indicate that such mutagenicity is likely, testing in an appropriate 5thstrain is not considered necessary. In addition, the only silicon-containing substance which has given a positive result in a bacterial strain capable of detecting cross-linking or oxidising mutagens contains an epoxy side-chain (which is associated with cross-linking mutagenicity), and this substance was positive in Salmonella typhimurium strains TA 100, TA 1535 as well as in E.coliWP2uvrA.
No measured data are available to assess the mutagenicity or cytogenicity potential of N-[3-(triethoxysilyl)propyl]ethylenediaminein mammalian cells, however, reliable data are available for the structural analogue substance, N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3).
READ-ACROSS JUSTIFICATION
To reduce animal testing REACH recommends to make use of a read-across approach where appropriate based on the high accordance in properties relevant for the specific endpoint. In the case of genetic toxicity relevant properties are structural similarity as well as physical-chemical and basic toxicological parameters in the same range. Especially functional groups that are associated with genetic toxicity have to be compared. In the following paragraphs the read-across approach for N-[3-(triethoxysilyl)propyl]ethylenediamine (CAS 5089-72-5) is evaluated point by point.
Read-across hypothesis
For N-[3-(triethoxysilyl)propyl]ethylenediamine an Ames test is available, which was negative in the Salmonella typhimurium strains TA 1535, TA 1537, TA 98, and TA 100 when tested up to the limit dose of 5000 µg/plate. No measured data are available to assess the mutagenicity or cytogenicity potential of N-[3-(triethoxysilyl)propyl]ethylenediamine in mammalian cells, however, reliable data are available for the structural analogue substance, N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3). None of these tests showed a positive result, indicating no genotoxic properties of the test substance. This is further supported by the fact that no positive results were observed with other substances in the amino functional alkoxysilanes analogue group (PFA, 2013t). The substances have the same or similar hydrolysis products and therefore read-across to the analogue is appropriate. The other hydrolysis product methanol is not genotoxic. Additional information is given in a supporting report attached in Section 13 of the IUCLID 5 dossier.
Discussion
Genetic toxicity (mutagenicity) in mammalian cells in vitro
N-(3-trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3) has been tested in a reliable study according to an appropriate US EPA test guideline that is equivalent to OECD 476 and under GLP (BRRC, 1988a). An increase in mutant frequency was observed in one of two duplicate cultures with metabolic activation at one concentration in the initial experiment. No other increase in mutant frequency was observed at any concentration (0.1 to 4.0 mg/ml) with and without activation (5 hours treatment). The experiment was repeated (with metabolic activation): no increase in mutant frequency was observed. Cytotoxicity was observed in all test strains at the top concentration of 4.0 mg/ml with metabolic activation. The appropriate solvent and positive controls were included and gave expected results. It is concluded that the test substance is negative for the induction of mutations in CHO cells under the conditions of the test.
Genetic toxicity (cytogenicity) in mammalian cells in vitro
No in vitro cytogenicity study is available; testing is not required as a reliable in vivo micronucleus assay is available for the structural analogue substance, N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3).
Genetic toxicity (mutagenicity) in mammalian cells in vivo
N-(3-(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3) has been tested in a reliable in vivo micronucleus study according to EPA Health Effect T G, Report 560/6-83-001, which is similar to OECD 474, and under GLP conditions (BRRC, 1988b). No treatment related increases in numbers of micronuclei in PCE's of Swiss-Webster mice were observed after intraperitoneal administration at concentrations of 87.5, 175 and 280 mg/kg bw. Relatively high dose levels of the test compound were tested up to 80% of the LD50 with no indication of significant induction of micronuclei. The PCE/NCE ratio was reduced at the highest concentration, 48 h exposure and at all doses, 72 h exposure, indicating exposure of the target tissue to the test substance. The test compound is considered to be negative for the induction of micronuclei under the conditions of this test.
In conclusion, no evidence of mutagenicity or clastogenicity was observed in any of the available in vitro and in vivo studies.
Justification for classification or non-classification
The available data on genetic toxicity of the analogue substance N-(3 -(trimethoxysilyl)propyl)ethylenediamine (CAS 1760-24-3) and the registered substance do not meet the criteria for classification according to Regulation (EC) No. 1272/2008 and are therefore conclusive but not sufficient for classification.
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.